Improving filterability of lactase by adding anion selected from malate, tartrate, citrate, gluconate, EDTA or combination thereof and obtained sterile filtered lactase product

Abstract

A sterile filtered liquid lactase composition comprising a lactase (preferably neutral or acidic lactase), an anion (selected from malate, tartrate, citrate, gluconate and / or EDTA) and a cation (selected from sodium and potassium). Preferably, the composition further comprises a polyol. Addition of the anion to lactase improves filterability by reducing pressure in the filtration system. Preferably, the filtration forms part of an in-line filtration system in the production of dairy products.

Description

Technical Field

[0001] The present invention generally relates to an improved formulation or improved composition of lactase or lactase products, wherein the formulation or composition is supplemented with an anion selected from malate, tartrate, citrate, gluconate and EDTA or a combination thereof. The formulation or composition helps improve the physical stability and filterability of lactase or lactase products.

[0002] background

[0003] At present, lactase or lactase products are only formulated with glycerol and tap water. Adding lactase or lactase products to milk or milk-based products can be implemented before or after the milk or milk-based products are sterilized. In the case of the latter, a step of sterilizing lactase by filtration is required. However, lactase or lactase products may easily cause filter clogging in the step of sterilizing lactase by filtration. This problem has been reported in multiple files such as EP 3 568 023 or EP 3 187 582, and causes the replacement of multiple filters, resulting in increased production costs and production delays.

[0004] Overview

[0005] It is an object of the present invention or disclosure to provide a formulation or composition of lactase or a lactase product that avoids filter clogging, or in other words, a formulation or composition of lactase or a lactase product with improved physical stability and / or filterability. This object is achieved when the formulation or composition is supplemented with anions selected from the list consisting of malate, tartrate, citrate, gluconate and ethylenediaminetetraacetate and combinations thereof, resulting in improved physical stability and / or filterability of the lactase formulation or composition or the lactase product.

[0006] In the context of the present invention or disclosure, when filtering a formulation or composition comprising lactase or a lactase product and an anion such as malate, tartrate, citrate, gluconate and ethylenediaminetetraacetate and / or combinations thereof, the improvement in physical stability and / or filterability is observed by a decrease in the pressure measured in the filtration system compared to a control. In the context of the present invention or disclosure, a control is defined as a formulation or composition comprising lactase or a lactase product but lacking a supplementary anion selected from malate, tartrate, citrate, gluconate and ethylenediaminetetraacetate and / or combinations thereof.

[0007] The present invention or disclosure indicates that anions selected from malate, tartrate, citrate, gluconate and / or combinations thereof help reduce pressure build-up in the filtration system and thereby allow longer filtration times before the pressure limit of the system is reached, thus preventing filter rupture. As a result, a larger volume of a preparation or composition comprising lactase or a lactase product and said anion can be filtered using the same filter before the filter needs to be changed, compared to the volume that can be filtered when filtering a preparation or composition comprising lactase or a lactase product or lactase lacking said anion.

[0008] None of the above documents mentions, suggests or indicates that the nature of the anion affects the filterability of a composition comprising lactase and said anion.

[0009] definition

[0010] For purposes of the present invention or disclosure, the following definitions apply.

[0011] As used herein, "neutral lactase" refers to a lactase having enzymatic activity, preferably optimal activity, at neutral pH.

[0012] As used herein, "acid lactase" refers to a lactase having enzymatic activity, preferably optimal activity, at acidic pH.

[0013] As used herein, "sequence identity" of amino acids refers to the sequence identity calculated as (n ref n dif )·100 / n ref The sequence identity of dif is the total number of non-identical residues in the two sequences when aligned, and where n refIs the number of residues in one of the sequences. In some embodiments, sequence identity is determined by, for example, Smith and Waterman, 1981, Adv. Appl. Math. 2: 482 conventional methods, by Pearson & Lipman, 1988, Proc. Natl. Acad. Sci. USA 85: 2444 similarity search methods, using Thompson et al., 1994, Nucleic Acids Res 22: 467380 CLUSTAL W algorithm, by computerized implementation of these algorithms (GAP, BESTFIT, FASTA and TFASTA in the Wisconsin Genetics software package of Genetics Computer Group). BLAST algorithm (Altschul et al., 1990, Mol. Biol. 215: 403-10) can also be used, and its software can be obtained through the National Center for Biotechnology Information (www.ncbi.nlm.nih.gov / ). When using any of the above algorithms, the default parameters for "window" length, gap penalties, etc. are used.

[0014] Composition and formulation are used synonymously herein.

[0015] Sodium citrate refers to trisodium citrate.

[0016] In the context of describing the present invention or disclosure (especially in the context of the following claims), the use of the terms "a, an" and "the" and similar references should be interpreted as covering the singular and plural, unless otherwise specified herein or clearly contradicted by the context. Unless otherwise specified, the terms "comprising", "having", "including" and "containing" should be interpreted as open terms (i.e., meaning "including, but not limited to"). Unless otherwise specified herein, the description of the numerical range herein is intended only as a shorthand method for referring to each individual numerical value within the range individually, and each individual value is incorporated into the specification as if it were described separately herein. Unless otherwise indicated herein or clearly contradicted by the context, all methods described herein can be performed in any suitable order. The use of any and all examples or exemplary language (e.g., "such as") provided herein is intended only to better illustrate the present invention or disclosure, rather than to limit the scope of the present invention or disclosure, unless otherwise required. Any language in the specification should not be interpreted as indicating that any unrequired element is essential to the practice of the present invention or disclosure.

[0017] Detailed description

[0018] The present invention or disclosure relates to a composition comprising lactase and an anion selected from malate, tartrate, citrate, gluconate and ethylenediaminetetraacetic acid and / or a combination thereof. As can be seen from the examples, the counterion (or cation in this case) can be selected from sodium and potassium. Preferably, the composition disclosed herein can include 0.1mM-250mM malate, 0.1mM-250mM tartrate, 0.1mM-250mM citrate, 0.1mM-250mM gluconate, 0.1mM-250mM ethylenediaminetetraacetic acid and / or a combination thereof. More preferably, the composition can include 1mM-150mM malate, 1mM-150mM tartrate, 1mM-150mM citrate, 1mM-150mM glucose salt sugar acid, 1mM-150mM ethylenediaminetetraacetic acid and / or a combination thereof. Even more preferably, the composition may comprise 2mM-100mM or 10mM-20mM malate, 2mM-100mM or 10mM-20mM tartrate, 2mM-100mM or 10mM-20mM citrate, 2mM-100mM or 10mM-20mM gluconate, 2mM-100mM or 10mM-20mM ethylenediaminetetraacetate and / or combinations thereof.

[0019] The present invention or the disclosure also relates to a composition comprising lactase and a salt, wherein the salt is selected from a list consisting of sodium malate, potassium malate, sodium tartrate, potassium tartrate, sodium citrate, potassium citrate, sodium gluconate, potassium gluconate, disodium ethylenediaminetetraacetic acid, dipotassium ethylenediaminetetraacetic acid, and / or a combination thereof. Preferably, the composition disclosed herein may comprise 0.1 mM-250 mM of sodium malate, potassium malate, sodium tartrate, potassium tartrate, sodium citrate, potassium citrate, sodium gluconate, potassium gluconate, disodium ethylenediaminetetraacetic acid, dipotassium ethylenediaminetetraacetic acid, and / or a combination thereof. More preferably, the composition may comprise 1 mM-150 mM of sodium malate, potassium malate, sodium tartrate, potassium tartrate, sodium citrate, potassium citrate, sodium gluconate, potassium gluconate, disodium ethylenediaminetetraacetic acid, dipotassium ethylenediaminetetraacetic acid, and / or a combination thereof. Even more preferably, the composition may include 2mM-100mM or 10mM-20mM sodium malate, potassium malate, sodium tartrate, potassium tartrate, sodium citrate, potassium citrate, sodium gluconate, potassium gluconate, disodium ethylenediaminetetraacetic acid, dipotassium ethylenediaminetetraacetic acid and / or combinations thereof.

[0020] Preferably, any composition disclosed herein may comprise between 20 BLU / g and 10000 BLU / g of lactase.

[0021] Preferably, any composition disclosed herein may comprise a neutral lactase or an acid lactase.

[0022] Preferably, the lactase may have an amino acid sequence that is at least 75%, or at least 80%, or at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 1-53. More preferably, the lactase may have an amino acid sequence that is 75%-100% or 80%-100% or 85%-100% identical to any one of SEQ ID NOs: 1-53, such as 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 1-53. Even more preferably, 90%-100% identical to any one of SEQ ID NO: 1-53, such as 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NO: 1-53. The most preferred sequences are SEQ ID NO: 1, 7, 40, 52, 53 and any sequence having 75%-100% or 80%-100% or 85%-100% identical to any one of SEQ ID NO: 1, 7, 40, 52, 53, such as 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NO: 1, 7, 40, 52, 53.

[0023] Any composition disclosed herein may also comprise a polyol selected from glycerol, sorbitol, monopropylene glycol, sucrose, glucose, galactose or lactose or a combination thereof. Preferably, the polyol may be present in a concentration of 10%-70% w / w, more preferably 20%-80% w / w, even more preferably 30%-60% w / w. Preferably, the polyol is or may be glycerol.

[0024] In addition, any composition disclosed herein may be a liquid lactase composition, preferably a sterile liquid lactase composition or a sterile filtered liquid lactase composition. The present invention or disclosure also relates to the use of anions selected from malate, tartrate, citrate, gluconate, ethylenediaminetetraacetate and / or combinations thereof to improve the filterability of any composition disclosed herein. As can be seen from the examples, the counterion (or cation in this case) may be sodium and / or potassium.

[0025] The present invention or disclosure also relates to the use of anions selected from malate, tartrate, citrate, gluconate, ethylenediaminetetraacetate and / or combinations thereof in reducing the pressure of a filtration system, preferably in reducing the pressure of an in-line filtration system in a dairy product production process.

[0026] Preferably, the use disclosed herein relates to 0.1mM-250mM malate, 0.1mM-250mM tartrate, 0.1mM-250mM citrate, 0.1mM-250mM gluconate, 0.1mM-250mM ethylenediaminetetraacetate and / or a combination thereof. More preferably, it relates to 1mM-150mM malate, 1mM-150mM tartrate, 1mM-150mM citrate, 1mM-150mM gluconate, 1mM-150mM ethylenediaminetetraacetate and / or a combination thereof. Even more preferably, it concerns 2mM-100mM or 10mM-20mM malate, 2mM-100mM or 10mM-20mM tartrate, 2mM-100mM or 10mM-20mM citrate, 2mM-100mM or 10mM-20mM gluconate, 2mM-100mM or 10mM-20mM ethylenediaminetetraacetate and / or combinations thereof.

[0027] The present invention or disclosure also relates to a method for reducing pressure in a filtration system, comprising the step of filtering any lactase composition disclosed herein. Preferably, wherein the filtration system is an online filtration system of a dairy product production process. More preferably, wherein the filtration system is a sterile online dosing system. Example

[0028] Any lactase or lactase product, in particular one that exhibits poor filterability, may be used in the following embodiments. For example, the lactase or lactase product may comprise a lactase from Bifidobacterium, Lactobacillus, Streptococcus or Kluyveromyces. Preferably, the lactase or lactase product may comprise an enzyme from Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium catenulatum, Bifidobacterium longum, Lactobacillus sakei, Lactobacillus amylovorus, Lactobacillus delbrueckii (such as Lactobacillus delbrueckii subsp. lactis or Lactobacillus delbrueckii subsp. bulgaricus or Lactobacillus delbrueckii subsp. indicus, Lactobacillus helvaticus, Lactobacillus reuteri, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus serrata, Lactobacillus reuteri, Lactobacillus crispatus, Streptococcus thermophilus, Kluyveromyces lactis.

[0029] More preferably, a lactase or lactase product represented by any of the sequences disclosed herein as SEQ ID NOs: 1 to 53 may be used in the present invention or disclosure. A lactase or lactase product having an amino acid sequence that is at least 75%, or at least 80%, or at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 1-53 may be used in the present invention or disclosure. More preferably, the lactase may have an amino acid sequence that is 75%-100% identical to any one of SEQ ID NOs: 1-53, or 80%-100% or 85%-100%, such as 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 1-53, which may be used in the present invention or disclosure. Even more preferably, 90%-100% identical to any one of SEQ ID NOs: 1-53, such as 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 1-53 may be used in the present invention or disclosure. The combination of lactase or lactase product with an anion such as malate, tartrate, citrate, gluconate, EDTA and / or combinations thereof results in a formulation or composition comprising the lactase or lactase product and the anion. As can be seen in the examples, the counterion (or cation in this case) may be sodium or potassium.

[0030] An example of a lactase from Bifidobacterium bifidum is Fit, Novozymes A / S or Nurica by IFF TM An example of a Kluyveromyces lactis lactase is Ha-Lactase from Chr. Hansen A / S. TM 、Novozymes A / S or DSM

[0031] An example of Lactobacillus delbrueckii subsp. bulgaricus is Bonlacta TM .

[0032] In the following examples, when filtering a control formulation or composition, the remaining pressure in the system was measured and normalized to 100%. In the same embodiment, when filtering a preparation or composition (also referred to as a test formulation or test composition) comprising lactase or a lactase product and an anion selected from malate, tartrate, citrate, gluconate and / or a combination thereof, the remaining pressure in the system was measured and normalized according to the control formulation used in the same embodiment. For completeness, the control formulation (or control composition) and the test formulation (or test composition) of the same volume were filtered. In addition, 2 independent measurements were performed for each sample.

[0033] Example 1

[0034] Will Fit 5500 batches (from Chr. Hansen A / S) were diluted into 50% (v / v) glycerol in tap water (control) or into 50% (v / v) glycerol in tap water supplemented with anions selected from malate, tartrate, citrate, gluconate, EDTA or a combination thereof to reach a final specification of 2750 BLU / g.

[0035] The resulting composition was pre-filtered through a 0.45 μm VWR media bottle top filter, PES 0.45 μm, 90 mm (Cat no#514-0300) and then stored at 5°C. After incubation at 37°C for 24 hours, the sample was placed at room temperature for 1 hour before the filterability test. The sample is preferably placed in a temperature range of 15°C to 30°C, preferably 15°C to 25°C, more preferably 20°C to 25°C (at 1 atm). The sample was filtered using a SartoFLOW smart filtration device through a 0.45 μm prefilter (Sartorius syringe filter, Minisart NML, CA, 28 mm, 0.45 μm, sterile, Cat#16555-K) and a 0.2 μm sterile filter (Pall Life Science Acrodisc 25 mm syringe filter 0,2 μm Fluorodyne II membrane (sterile) VWR cat#516-8973P) and pre-wetted with MilliQ water (ELGA Purelab flex). The flow rate was stabilized at 6.5 g / min (+ / -10%), and the initial rate of increase of the system back pressure was measured and calculated by linear fitting in bar / hour.

[0036] The increase in system pressure rate was used to assess the filterability of the sample, meaning that samples that resulted in a higher pressure increase rate had lower filterability.

[0037] Table 1. Sample 1 was subjected to a 24 h stress period at 37°C.

[0038]

[0039] Table 1 shows that citrate, especially sodium citrate, performs better than the control or chloride, especially sodium chloride, in reducing the pressure generated when filtering the lactase composition. As a result, a higher volume of the lactase composition comprising citrate can be filtered before the filter coagulates compared to the volume of the control lactase composition or the control lactase composition comprising sodium chloride.

[0040] In addition, Table 1 shows that lower concentrations of citrate (particularly sodium citrate) are required to promote pressure reduction in the filtration system compared to the concentrations required when using chloride or sodium chloride. For example, 10 mM sodium citrate (about 0.3% W 柠檬酸钠 / W 乳糖酶成分 ) resulted in a significant decrease in the measured pressure, whereas 200 mM NaCl (approximately 1% W 氯化钠 / W 乳糖酶组合物 ) has a much smaller effect on reducing pressure.

[0041] Therefore, citrate is a better solution than chloride to improve the filterability of a formulation or composition comprising lactase or a lactase product. Preferably, sodium citrate is a better solution than sodium chloride. Furthermore, similar results are expected for any lactase, in particular any lactase from bifidobacteria, including the lactase from bifidobacteria disclosed herein.

[0042] Example 2

[0043] Example 2 was carried out as in Example 1, except that several concentrations of sodium citrate were tested - Table 2.

[0044] Table 2. Sample 2 was subjected to a 24 h stress period at 37°C.

[0045]

[0046]

[0047] Table 2 shows that regardless of the concentration of citrate (preferably sodium citrate is used), a reduction in pressure rate is measured when filtering a composition comprising lactase and sodium citrate. Furthermore, the results obtained with 10 mM sodium citrate shown in Tables 1 and 2 are consistent, both showing that the pressure remaining in the filtration is about 41% to 49%. Therefore, the pressure in the system is reduced by more than 50% compared to the control, indicating that the different samples of lactase or lactase product show improved filterability of the lactase or lactase product. Furthermore, similar results are expected for any lactase, particularly any lactase from bifidobacteria, including the lactase from bifidobacteria disclosed herein.

[0048] Example 3

[0049] Example 3 was carried out as in Example 1, except that different anions were tested - Table 3.

[0050] Table 3. Samples 2 and 3 were subjected to a stress period of 24 h at 37 °C.

[0051]

[0052] Table 3 confirms the results obtained with 10 mM sodium citrate and as shown in Tables 1 and 2. In other words, when 10 mM sodium citrate is used, it can reduce the pressure of the filtration system by more than 50%. This observation was consistent using 3 different samples, thus indicating that the effect is independent of the sample.

[0053] Furthermore, a reduction in pressure (and therefore improved filterability) was also observed when potassium citrate was used instead of sodium citrate, illustrating the effect of citrate in improving the filterability of lactase or lactase products. Preferably, 10 mM potassium citrate can reduce pressure and therefore improve filterability similarly to 10 mM sodium citrate. Thus, Table 3 shows the effect of citrate in improving filterability regardless of which cation (potassium or sodium) is used.

[0054] Table 3 also shows the pressure improvement when anions other than citrate are used in the formulation or composition. The improvement is exemplified when gluconate, tartrate, malate or edetate is added to the lactase or lactase product. Preferably, the improvement is exemplified by gluconate. More preferably, the improvement is exemplified by tartrate or malate (which results in a pressure reduction of about 50%). Even more preferably, the improvement is exemplified by edetate. When edetate is used, the remaining pressure in the filtration system is about 29%, which means that the pressure drops by about 71%. All values ​​are given compared to the control. Preferably, the improvement is exemplified by potassium gluconate. More preferably, the improvement is exemplified by sodium tartrate or sodium malate. Even more preferably, the improvement is exemplified by disodium edetate. All values ​​are given compared to the control.

[0055] Thus, the effects of citrate observed in Tables 1 to 3 can be extended to other anions, such as malate, tartrate, gluconate or EDTA. Preferably, the filterability improvement extends to potassium citrate, sodium tartrate, sodium malate, potassium gluconate or disodium EDTA, regardless of the lactase sample tested. Furthermore, similar results are expected for any lactase, particularly any lactase from bifidobacteria, including the lactase from bifidobacteria disclosed herein.

[0056] Example 4

[0057] Example 4 was carried out as in Example 1, except that the lactase prototype was obtained from Chr. Hansen A / S. Fit non-standardized samples were prepared and diluted in 50% glycerol (+ sodium citrate) in tap water to 5500 BLU / g strength or 2100 BLU / g strength. Lactase was exposed to a 6-week stress period at 5°C (Table 4) or 6 weeks at 5°C followed by 24 hours incubation at 37°C (Table 5). The 6-week stress period at 5°C is one of the possible representative forms of storage conditions.

[0058] Table 4. Sample 4 was subjected to a stress period of 6 weeks at 5°C.

[0059]

[0060] Table 5. Sample 4 was subjected to a stress period of 6 weeks at 5°C and then 24 hours at 37°C before filtration.

[0061]

[0062] Both tables (Table 4 and Table 5) show the relevance of supplementing a formulation or composition comprising lactase or a lactase product with an anion such as citrate, which is then placed under typical storage conditions. Even after the storage period, when citrate is used or supplemented in a formulation or composition comprising lactase or a lactase product, the formulation of the composition maintains improved filterability characteristics because the remaining pressure in the system remains lower than the pressure of the control. In addition, the observed effect is independent of the enzyme dosage or specification used because the pressure retained in the system is smaller for both dosages. All values ​​are given in comparison with the control. Thus, Tables 4 and 5 show that the effect of anions is not time-limited or independent of enzyme specification. In addition, the remaining anions (malate, tartrate, gluconate, ethylenediaminetetraacetic acid and / or combinations thereof) are also expected (based on the previous examples) to exhibit this effect. Finally, similar results are expected for any lactase, in particular any lactase from bifidobacteria, including lactases from bifidobacteria disclosed herein.

[0063] Example 5

[0064] Lactase from the genus Bifidobacterium was also tested, in particular SEQ ID NO: 40-Sample 5 from Bifidobacterium longum. The non-standardized concentrated sample was diluted to 14000 BLU / g specification in 50% glycerol (+ sodium citrate or sodium tartrate) in tap water. In this example, the specification of the lactase used was determined by comparing the activity of the lactase with the activity of a lactase with a known specification, which can be, for example, the same lactase used in Examples 1 to 4. Alternatively, the specification can also be determined by multiplying by a factor of 2 as described in Examples 16 and 17 of WO2018189224 or as described in EP 3 568 023. The resulting 14000 prototype was pre-filtered through a 0.45 μm VWR media bottle top filter, PES 0.45 μm, 90 mm (Cat no#514-0300) and stored at 5°C. The sample was placed at room temperature for 1 h and then subjected to a filterability test. The sample was filtered using a SartoFLOW smart filtration device, sequentially through a 0.45 μm prefilter (Sartorius syringe filter, Minisart NML, CA, 28 mm, 0.45 μm, sterile, Cat#16555-K) and a 0.2 μm sterile filter (Pall LifeScience Acrodisc 25 mm syringe filter 0.2 μm Fluorodyne II membrane (sterile) (VWR cat: 516-8973P), and pre-wetted with MilliQ water (ELGA Purelab flex). The flow rate was stabilized at 6.5 g / min (+ / -10%), and the initial rate of increase of the system back pressure was measured and calculated by linear fitting in bar / hour. The increase in back pressure rate is used to evaluate the filterability of the sample, which means that samples that produce a higher pressure increase rate have lower filterability.

[0065] Table 6. Sample 5 was subjected to a stress period of 4 days at 5°C.

[0066]

[0067] Table 6 shows that the pressure is reduced when filtering sample 5, and thus shows that the lactase sample filterability is improved when using citrate or tartrate. In addition, the combination of Example 5 with the remaining examples disclosed herein shows that the pressure of the filtration system is reduced when malate, tartrate, citrate, gluconate, ethylenediaminetetraacetic acid or a combination thereof is used in the formulation or composition of lactase or lactase product (regardless of which lactase is used). Therefore, Example 5 confirms that similar results are obtained regardless of which lactase or lactase product is used, preferably, for any lactase from bifidobacteria, including lactase from bifidobacteria disclosed herein, similar results are obtained, more preferably, for any lactase from Bifidobacterium bifidum or Bifidobacterium longum, similar results are obtained, as exemplified in Examples 1 to 5.

[0068] Example 6

[0069] Also tested was a lactase from Kluyveromyces, specifically SEQ ID NO: 53-Sample 6 from Kluyveromyces lactis. This lactase is also known as HA- HA 5200 batches (from Chr. Hansen A / S) were diluted into 50% (v / v) glycerol in tap water (control) or supplemented with anions selected from tartrate and citrate or a combination thereof to reach a final specification of 2600 BLU / g. In this example, the specification of the lactase used is determined by comparing the activity of the lactase with the activity of a lactase with a known specification, which can be, for example, the same lactase used in Examples 1 to 4. Alternatively, the specification can also be determined as described in Examples 16 and 17 of WO2018189224. The resulting 2600 prototype was pre-filtered through a 0.45μm VWR media bottle top filter unit, PES 0.45μm, 90mm (Cat no#514-0300) and stored at 5°C. After incubation at 37°C for 24h, the sample was incubated at room temperature for 1h and then subjected to filterability testing. The sample is preferably placed in a temperature range of 15°C to 30°C, preferably 15°C to 25°C, more preferably 20°C to 25°C (at 1 atm). Using a SartoFLOW intelligent filtration device, the sample is filtered sequentially through a 0.45 μm pre-filter (Sartorius syringe filter, Minisart NML, CA, 28 mm, 0.45 μm, sterile, Cat#16555-K) and a 0.2 μm sterile filter (Pall Life Science Acrodisc 25 mm syringe filter 0,2 μm Fluorodyne II membrane (sterile) (VWR cat: 516-8973P) and pre-wetted with MilliQ water (ELGA Purelab flex). The flow rate is stabilized at 6.5 g / min (+ / -10%), and the initial rate of increase of the system back pressure is measured and calculated by linear fitting in bar / hour. The increase in back pressure rate is used to evaluate the filterability of the sample, which means that samples that produce a higher pressure increase rate have lower filterability.

[0070] Table 7. Sample 6 was subjected to a 24 h stress period at 37°C.

[0071]

[0072] Table 7 shows that the pressure was reduced when filtering sample 6, thus showing the improvement in the filterability of the lactase sample when using citrate or tartrate. In addition, Example 6 in combination with the remaining examples disclosed herein shows that the pressure of the filtration system is reduced when malate, tartrate, citrate, gluconate, EDTA or a combination thereof is used in the formulation or composition of lactase or lactase product (regardless of which lactase is used). Thus, Example 6 demonstrates that similar results are obtained regardless of which lactase or lactase product is used, preferably for any lactase from Bifidobacterium or Kluyveromyces, including lactases from these two genera disclosed herein, and more preferably for any lactase from Bifidobacterium bifidum or Bifidobacterium longum or Kluyveromyces lactis.

[0073] In summary, the filterability of a lactase sample may be improved if malate, tartrate, citrate, gluconate, EDTA or a combination thereof is part of a preparation or composition comprising lactase or a lactase product prior to the filtration step. The cation may be sodium or potassium.

[0074] Sequence Listing

[0075]

[0076]

[0077]

[0078]

[0079]

[0080]

[0081]

[0082]

[0083]

[0084]

[0085]

[0086] Numbered implementation plan

[0087] Composition

[0088] Implementation 1.A composition comprising lactase and an anion selected from malate, tartrate, citrate, gluconate, ethylenediaminetetraacetate and / or combinations thereof.

[0089] Implementation 2. A composition according to the aforementioned embodiment, wherein the anion is 0.1 mM to 250 mM malate, tartrate, citrate, gluconate, ethylenediaminetetraacetate and / or a combination thereof; preferably 1 mM to 150 mM malate, tartrate, citrate, gluconate, ethylenediaminetetraacetate and / or a combination thereof; more preferably 2 mM to 100 mM malate, tartrate, citrate, gluconate, ethylenediaminetetraacetate and / or a combination thereof; even more preferably 10 mM to 20 mM malate, tartrate, citrate, gluconate, ethylenediaminetetraacetate and / or a combination thereof.

[0090] Implementation 3. The composition according to any of the preceding embodiments 1-2, wherein the lactase is neutral lactase or acid lactase.

[0091] Implementation 4.A composition according to any of the preceding embodiments 1-3, wherein the lactase has an amino acid sequence that is at least 75% identical to any one of SEQ ID NOs: 1-53, preferably at least 80%, or at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 1-53. More preferably, the lactase may have an amino acid sequence that is 75%-100% identical to any one of SEQ ID NOs: 1-53, or 80%-100% or 85%-100% identical to any one of SEQ ID NOs: 1-53, such as 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 1-53. Even more preferably, 90%-100% identical to any one of SEQ ID NO: 1-53, such as 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NO: 1-53. The most preferred sequences are SEQ ID NO: 1, 7, 40, 52, 53 and any sequence having 75%-100% or 80%-100% or 85%-100% identical to any one of SEQ ID NO: 1, 7, 40, 52, 53, such as 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NO: 1, 7, 40, 52, 53.

[0092] Implementation 5. The composition according to any one of the preceding embodiments 1-4, comprising a polyol selected from glycerol, sorbitol, monopropylene glycol, sucrose, glucose, galactose or lactose or a combination thereof, preferably glycerol.

[0093] Implementation 6. According to the composition of the aforementioned embodiment 5, the concentration of the polyol is 10%-70%. 多元醇 / W 组合物 , preferably 20%-80% W 多元醇 / W 组合物 , more preferably 30%-60% W 多元醇 / W 组合物 .

[0094] Implementation Plan 7. The composition according to any one of the preceding embodiments 1-6, wherein the composition is a liquid lactase composition.

[0095] Implementation 8. The composition according to any one of the preceding embodiments 1-7, wherein the composition is a sterile liquid lactase composition.

[0096] Embodiment 9. A sterile filtered liquid lactase composition comprising:

[0097] Lactase;

[0098] an anion selected from the list consisting of malate, tartrate, citrate, gluconate, and ethylenediaminetetraacetate;

[0099] And preferably has a cation selected from the list consisting of sodium and potassium.

[0100] Embodiment 10. A sterile filtered liquid lactase composition according to embodiment 9, wherein the anion is selected from the list consisting of 0.1 mM to 250 mM malate, tartrate, citrate, gluconate and EDTA, preferably 1 mM to 150 mM malate, tartrate, citrate, gluconate and EDTA, more preferably 2 mM to 100 mM malate, tartrate, citrate, gluconate and EDTA, even more preferably 10 mM to 20 mM malate, tartrate, citrate, gluconate and EDTA.

[0101] Embodiment 11. A sterile filtered liquid lactase composition according to any one of the preceding embodiments 9-11, wherein the anion and the cation form a salt, and the salt is selected from the list consisting of sodium malate, potassium malate, sodium tartrate, potassium tartrate, sodium citrate, potassium citrate, sodium gluconate, potassium gluconate, disodium ethylenediaminetetraacetic acid, and dipotassium ethylenediaminetetraacetic acid; preferably, wherein the salt is selected from the list consisting of 0.1 mM-250 mM sodium malate, potassium malate, sodium tartrate, potassium tartrate, sodium citrate, potassium citrate, sodium gluconate, potassium gluconate, disodium ethylenediaminetetraacetic acid, and dipotassium ethylenediaminetetraacetic acid, more preferably 1 mM. 2mM-150mM sodium malate, potassium malate, sodium tartrate, potassium tartrate, sodium citrate, potassium citrate, sodium gluconate, potassium gluconate, disodium ethylenediaminetetraacetic acid, and dipotassium ethylenediaminetetraacetic acid, even more preferably a list consisting of 2mM-100mM sodium malate, potassium malate, sodium tartrate, potassium tartrate, sodium citrate, potassium citrate, sodium gluconate, potassium gluconate, disodium ethylenediaminetetraacetic acid, and dipotassium ethylenediaminetetraacetic acid, or 10mM-20mM sodium malate, potassium malate, sodium tartrate, potassium tartrate, sodium citrate, potassium citrate, sodium gluconate, potassium gluconate, disodium ethylenediaminetetraacetic acid, and dipotassium ethylenediaminetetraacetic acid.

[0102] Embodiment 12. The sterile filtered liquid lactase composition according to any one of the preceding embodiments 9-11, wherein the lactase is a neutral lactase or an acid lactase.

[0103] Embodiment 13. A sterile filtered liquid lactase composition according to any one of embodiments 9-12, wherein the lactase has an amino acid sequence that is at least 75% identical to any one of SEQ ID NOs: 1-53, preferably 85% identical to any one of SEQ ID NOs: 1-53, more preferably 90% identical to any one of SEQ ID NOs: 1-53, preferably at least 80%, or at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 1-53. More preferably, the lactase may have an amino acid sequence that is 75%-100% identical to any one of SEQ ID NOs: 1-53, or 80%-100% or 85%-100% identical, such as 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 1-53. Even more preferably, it is 90%-100% identical to any one of SEQ ID NOs: 1-53, such as 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 1-53. The most preferred sequences are SEQ ID NO: 1, 7, 40, 52, 53 and any sequence that is 75%-100% or 80%-100% or 85%-100% identical to any one of SEQ ID NO: 1, 7, 40, 52, 53, for example, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NO: 1, 7, 40, 52, 53.

[0104] Embodiment 14. The sterile filtered liquid lactase composition according to any one of embodiments 9 to 13, comprising a polyol selected from glycerol, sorbitol, monopropylene glycol, sucrose, glucose, galactose or lactose or a combination thereof, preferably glycerol.

[0105] Embodiment 15. A sterile filtered liquid lactase composition according to embodiment 14, wherein the concentration of the polyol is 10%-70%. 多元醇 / W 组合物 , preferably 20%-80% W 多元醇 / W组合物 , more preferably 30%-60% W 多元醇 / W 组合物 .

[0106] use

[0107] Implementation Plan 16. Use of an anion selected from malate, tartrate, citrate, gluconate, ethylenediaminetetraacetate and / or a combination thereof for improving the filterability of the composition according to any one of the preceding embodiments 1 to 8. Thus, the anion is used as an improver of the filterability of the composition disclosed herein.

[0108] Implementation Plan 17. Use of anions selected from malate, tartrate, citrate, gluconate, ethylenediaminetetraacetate and / or combinations thereof for reducing the pressure of a filtration system of a composition according to any one of the preceding embodiments 1 to 8. Thus, the anions are used as a decompression element or agent of a filtration system for filtering the composition disclosed herein.

[0109] Implementation 18. When the composition as disclosed herein (any of the aforementioned embodiments 1-8) is used in the production process of dairy products, anions selected from malate, tartrate, citrate, gluconate, ethylenediaminetetraacetic acid and / or combinations thereof are used to reduce the pressure of the online filtration system of the dairy production process. Therefore, the anions are used as a decompression element or agent of the online filtration system, which is used to filter the composition disclosed herein.

[0110] Implementation Plan 19. The use according to any of the preceding embodiments 16-18, wherein the anion is 0.1 mM to 250 mM malate, tartrate, citrate, gluconate, ethylenediaminetetraacetate and / or a combination thereof; preferably 1 mM to 150 mM malate, tartrate, citrate, gluconate, ethylenediaminetetraacetate and / or a combination thereof; more preferably 2 mM to 100 mM malate, tartrate, citrate, gluconate, ethylenediaminetetraacetate and / or a combination thereof; even more preferably 10 mM to 20 mM malate, tartrate, citrate, gluconate, ethylenediaminetetraacetate and / or a combination thereof.

[0111] Embodiment 20. Anions selected from the list consisting of malate, tartrate, citrate, gluconate and ethylenediaminetetraacetic acid are used to improve the filterability of a composition, and / or to reduce the pressure of a filtration system of a composition, preferably for reducing the pressure of an online filtration system of a dairy production process, wherein the composition comprises lactase and the anions. Thus, the anions are used as filterability improvers of the compositions disclosed herein, and / or the anions are used as decompression elements or agents of a filtration system, which are used to filter the compositions disclosed herein, preferably the anions are used as decompression elements or agents of an online filtration system, which are used to filter the compositions disclosed herein.

[0112] Embodiment 21. The use according to embodiment 20, wherein the anion is selected from the list consisting of 0.1 mM to 250 mM malate, tartrate, citrate, gluconate and ethylenediaminetetraacetate, preferably 1 mM to 150 mM malate, tartrate, citrate, gluconate and ethylenediaminetetraacetate, more preferably 2 mM to 100 mM malate, tartrate, citrate, gluconate and ethylenediaminetetraacetate, even more preferably 10 mM to 20 mM malate, tartrate, citrate, gluconate and ethylenediaminetetraacetate.

[0113] Embodiment 22. The use according to any one of embodiments 20-21, wherein the composition comprises a cation selected from the list consisting of sodium and potassium.

[0114] Embodiment 23. The use according to any one of embodiments 20-22, wherein the anion and the cation form a salt selected from the list consisting of sodium malate, potassium malate, sodium tartrate, potassium tartrate, sodium citrate, potassium citrate, sodium gluconate, potassium gluconate, disodium ethylenediaminetetraacetic acid, and dipotassium ethylenediaminetetraacetic acid.

[0115] Embodiment 24. The use according to any one of the preceding embodiments 20-23, wherein the salt is selected from 1 mM-250 mM sodium malate, potassium malate, sodium tartrate, potassium tartrate, sodium citrate, potassium citrate, sodium gluconate, potassium gluconate, ethylenediaminetetraacetic acid disodium salt and ethylenediaminetetraacetic acid dipotassium salt, preferably 1 mM-150 mM sodium malate, potassium malate, sodium tartrate, potassium tartrate, sodium citrate, potassium citrate, sodium gluconate, potassium gluconate, ethylenediaminetetraacetic acid Disodium salt and dipotassium salt of EDTA, more preferably 2mM-100mM sodium malate, potassium malate, sodium tartrate, potassium tartrate, sodium citrate, potassium citrate, sodium gluconate, potassium gluconate, disodium salt of EDTA and dipotassium salt of EDTA, even more preferably 10mM-20mM list consisting of sodium malate, potassium malate, sodium tartrate, potassium tartrate, sodium citrate, potassium citrate, sodium gluconate, potassium gluconate, disodium salt of EDTA and dipotassium salt of EDTA.

[0116] Embodiment 25. The use according to any one of embodiments 20-24, wherein the lactase is neutral lactase or acid lactase.

[0117] Embodiment 26. The use according to any one of embodiments 20-25, wherein the lactase has an amino acid sequence that is at least 75% identical to any one of SEQ ID NOs: 1-53, preferably at least 85% identical to any one of SEQ ID NOs: 1-53, more preferably at least 90% identical to any one of SEQ ID NOs: 1-53, preferably at least 80%, or at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 1-53. More preferably, the lactase may have an amino acid sequence that is 75%-100% identical to any one of SEQ ID NOs: 1-53, or 80%-100% or 85%-100% identical, such as 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 1-53. Even more preferably, 90%-100% identical to any one of SEQ ID NOs: 1-53, such as 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 1-53. The most preferred sequences are SEQ ID NO: 1, 7, 40, 52, 53 and any sequence that is 75%-100% or 80%-100% or 85%-100% identical to any one of SEQ ID NO: 1, 7, 40, 52, 53, for example, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NO: 1, 7, 40, 52, 53.

[0118] Embodiment 27. The use according to any one of embodiments 20-26, wherein the composition comprises a polyol selected from glycerol, sorbitol, monopropylene glycol, sucrose, glucose, galactose or lactose or a combination thereof, preferably glycerol; more preferably wherein the concentration of the polyol is 10%-70%. 多元醇 / W 组合物 , preferably 20%-80% W 多元醇 / W 组合物 , more preferably 30%-60% W 多元醇 / W 组合物 .

[0119] Embodiment 28. The use according to any one of embodiments 20-27, wherein the composition is a liquid lactase composition or a sterile liquid lactase composition or a sterile filtered liquid lactase composition.

[0120] method

[0121] Implementation Plan 29. A method for reducing pressure in a filtration system comprising the step of filtering the lactase composition according to any one of the preceding embodiments 1-8.

[0122] Implementation Plan 30. A method according to the aforementioned embodiment 29, wherein the filtration system is an online filtration system of a dairy product production process.

[0123] Implementation Plan 31. A method according to any of the preceding embodiments 29-30, wherein the filtration system is a sterile in-line quantitative system.

[0124] Embodiment 32. A method of reducing pressure in a filtration system, comprising the step of filtering a composition in the filtration system, wherein the composition comprises lactase and an anion selected from the list consisting of malate, tartrate, citrate, gluconate, and ethylenediaminetetraacetate.

[0125] Embodiment 33. A method according to embodiment 32, wherein the anion is selected from the list consisting of 0.1 mM to 250 mM malate, tartrate, citrate, gluconate, ethylenediaminetetraacetate and / or combinations thereof; preferably 1 mM to 150 mM malate, tartrate, citrate, gluconate, ethylenediaminetetraacetate and / or combinations thereof; more preferably 2 mM to 100 mM malate, tartrate, citrate, gluconate, ethylenediaminetetraacetate and / or combinations thereof, even more preferably 10 mM to 20 mM malate, tartrate, citrate, gluconate, ethylenediaminetetraacetate and / or combinations thereof.

[0126] Embodiment 34. The method of any one of Embodiments 32-33, comprising a cation selected from sodium and potassium.

[0127] Embodiment 35. A method according to any one of embodiments 32-34, wherein the anion and the cation form a salt selected from the list consisting of sodium malate, potassium malate, sodium tartrate, potassium tartrate, sodium citrate, potassium citrate, sodium gluconate, potassium gluconate, disodium ethylenediaminetetraacetic acid, and dipotassium ethylenediaminetetraacetic acid.

[0128] Embodiment 36. A method according to any one of the preceding embodiments 32-35, wherein the salt is selected from the list consisting of 0.1 mM-250 mM, preferably 1 mM-150 mM, more preferably 2 mM-100 mM, and even more preferably 10 mM-20 mM sodium malate, potassium malate, sodium tartrate, potassium tartrate, sodium citrate, potassium citrate, sodium gluconate, potassium gluconate, disodium ethylenediaminetetraacetic acid, and dipotassium ethylenediaminetetraacetic acid.

[0129] Embodiment 37. The method of any one of embodiments 32-36, wherein the lactase is neutral lactase or acid lactase.

[0130] Embodiment 38. A method according to any one of embodiments 32-37, wherein the lactase has an amino acid sequence that is at least 75% identical to any one of SEQ ID NOs: 1-53, preferably at least 85% identical to any one of SEQ ID NOs: 1-53, more preferably at least 90% identical to any one of SEQ ID NOs: 1-53, preferably at least 80%, or at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 1-53. More preferably, the lactase may have an amino acid sequence that is 75%-100% identical to any one of SEQ ID NOs: 1-53, or 80%-100% or 85%-100% identical, such as 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 1-53. Even more preferably, 90%-100% identical to any one of SEQ ID NOs: 1-53, such as 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 1-53. The most preferred sequences are SEQ ID NO: 1, 7, 40, 52, 53 and any sequence that is 75%-100% or 80%-100% or 85%-100% identical to any one of SEQ ID NO: 1, 7, 40, 52, 53, for example, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NO: 1, 7, 40, 52, 53.

[0131] Embodiment 39. The method according to any one of embodiments 32-38, wherein the composition comprises a polyol selected from glycerol, sorbitol, monopropylene glycol, sucrose, glucose, galactose or lactose or a combination thereof, preferably glycerol; more preferably wherein the concentration of the polyol is 10%-70%. 多元醇 / W 组合物 , preferably 20%-80% W 多元醇 / W 组合物 , more preferably 30%-60% W 多元醇 / W 组合物 .

[0132] Embodiment 40. The method of any one of embodiments 32-39, wherein the composition is a liquid lactase composition or a sterile liquid lactase composition or a sterile filtered liquid lactase composition.

[0133] Embodiment 41. The method of any one of Embodiments 32-40, wherein the filtration system is an in-line filtration system of a dairy production process.

[0134] Embodiment 42. The method of any one of Embodiments 32-41, wherein the filtration system is a sterile, in-line, quantitative system.

Claims

1. A sterile filtered liquid lactase composition comprising: Lactase; an anion selected from the list consisting of malate, tartrate, citrate, gluconate, and ethylenediaminetetraacetate; and A cation selected from the list consisting of sodium and potassium.

2. The sterile filtered liquid lactase composition according to claim 1, wherein the anion is selected from the list consisting of malate, tartrate, citrate, gluconate and EDTA in an amount of 0.1 mM to 250 mM, preferably 1 mM to 150 mM, more preferably 2 mM to 100 mM, even more preferably 10 mM to 20 mM.

3. The sterile filtered liquid lactase composition according to any one of claims 1-2, wherein the lactase is a neutral lactase or an acid lactase and / or has an amino acid sequence that is at least 75% identical to any one of SEQ ID NOs: 1-53, preferably at least 85% identical to any one of SEQ ID NOs: 1-53, more preferably at least 90% identical to any one of SEQ ID NOs: 1-53.

4. The sterile filtered liquid lactase composition according to any one of claims 1 to 3, comprising a polyol selected from glycerol, sorbitol, monopropylene glycol, sucrose, glucose, galactose, lactose and / or a combination thereof, preferably glycerol; more preferably wherein the concentration of the polyol is 10%-70% W 多元醇 / W 组合物 , preferably 20%-80%W 多元醇 / W 组合物 , more preferably 30%-60%W 多元醇 / W 组合 things.

5. Use of an anion selected from the list consisting of malate, tartrate, citrate, gluconate and ethylenediaminetetraacetate for improving the filterability of a composition, and / or for reducing the pressure of a filtration system of a composition, preferably for reducing the pressure of an online filtration system of a dairy production process, wherein the composition comprises lactase and the anion.

6. Use according to claim 5, wherein the anion is selected from the list consisting of malate, tartrate, citrate, gluconate and ethylenediaminetetraacetate in an amount of 0.1 mM to 250 mM, preferably 1 mM to 150 mM, more preferably 2 mM to 100 mM, even more preferably 10 mM to 20 mM.

7. Use according to any one of claims 5-6, wherein the composition comprises a cation selected from the list consisting of sodium and potassium.

8. The use according to any one of claims 5-7, wherein the lactase is a neutral lactase or an acid lactase and / or has an amino acid sequence that is at least 75% identical to any one of SEQ ID NOs: 1-53, preferably at least 85% identical to any one of SEQ ID NOs: 1-53, more preferably at least 90% identical to any one of SEQ ID NOs: 1-53.

9. The use according to any one of claims 5 to 8, wherein the composition comprises a polyol selected from glycerol, sorbitol, monopropylene glycol, sucrose, glucose, galactose or lactose or a combination thereof, preferably glycerol; more preferably wherein the concentration of the polyol is 10% to 70%. 多元醇 / W 组合物 , preferably 20%-80%W 多元醇 / W 组合物 , more preferably 30%-60%W 多元醇 / W 组合物 .

10. The use according to any one of claims 5 to 9, wherein the composition is a liquid lactase composition or a sterile liquid lactase composition or a sterile filtered liquid lactase composition.

11. A method of reducing pressure in a filtration system comprising the step of filtering a composition in the filtration system, wherein the composition comprises lactase and an anion selected from the list consisting of malate, tartrate, citrate, gluconate and ethylenediaminetetraacetate.

12. The method according to claim 11, wherein the anion is selected from the list consisting of malate, tartrate, citrate, gluconate and ethylenediaminetetraacetate in an amount of 0.1 mM to 250 mM, preferably 1 mM to 150 mM, more preferably 2 mM to 100 mM, even more preferably 10 mM to 20 mM.

13. The method according to any one of claims 11-12, comprising a cation selected from the list consisting of sodium and potassium.

14. The method according to any one of claims 11-13, wherein the lactase is a neutral lactase or an acid lactase and / or has an amino acid sequence that is at least 75% identical to any one of SEQ ID NOs: 1-53, preferably at least 85% identical to any one of SEQ ID NOs: 1-53, more preferably at least 90% identical to any one of SEQ ID NOs: 1-53.

15. The method according to any one of claims 11 to 14, wherein the composition comprises a polyol selected from glycerol, sorbitol, monopropylene glycol, sucrose, glucose, galactose or lactose or a combination thereof, preferably glycerol; more preferably wherein the concentration of the polyol is 10% to 70% W 多元醇 / W 组合物 , preferably 20%-80%W 多元醇 / W 组合物 , more preferably 30%-60%W 多元醇 / W 组合物 .

16. The method according to any one of claims 11-15, wherein the composition is a liquid lactase composition or a sterile liquid lactase composition or a sterile filtered liquid lactase composition.

17. The method according to any one of claims 11-16, wherein the filtration system is an in-line filtration system of a dairy production process.

18. The method according to any one of claims 11-17, wherein the filtration system is a sterile in-line quantitative system.

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