Methods for extracting β-nicotinamide mononucleotide from microalgae and microalgae extracts

Abstract

This disclosure relates to a method for extracting β-nicotinamide mononucleotide from microalgae and the microalgae extract. The method includes: mixing microalgae raw material with an extractant for extraction; wherein the microalgae raw material is selected from one or more of Chlorella, Spirulina, and Haematococcus pluvialis; the extractant comprises 0.05–1.5 wt% of a nonionic surfactant, 0.5–1.0 wt% of an organic acid, 20–40 wt% of an alcohol, and the balance being a solvent. Microalgae raw materials are easy to cultivate, grow rapidly, have high protein content, and high β-nicotinamide mononucleotide content; the extraction method disclosed herein is simple, has a high extraction rate, and is low in cost.

Description

Technical Field

[0001] This disclosure relates to the field of natural microbial extraction technology, and more specifically, to a method for extracting β-nicotinamide mononucleotide from microalgae and a microalgae extract. Background Technology

[0002] β-Nicotinamide mononucleotide (NMN) is a product of the nicotinamide phosphoribosyltransferase reaction and a key precursor in NAD+ synthesis. NAD+ is a crucial coenzyme for cellular energy metabolism and oxidative stress adaptation. The NAD+-centric theory of mammalian aging posits that NAD+ levels determine the rate and extent of aging. NAD+ is present in all human cells and serves as an important cofactor or co-substrate, involved in various enzymatic processes, including glycolysis, the TCA cycle, oxidative phosphorylation, DNA repair, and protein deacetylation. NAD+ levels are vital for maintaining mitochondrial homeostasis, the body's metabolism, normal organ and tissue function, and delaying aging.

[0003] Currently, NMN is mainly prepared through chemical synthesis, enzymatic synthesis, and microbial fermentation. However, it faces challenges such as difficult synthesis and high costs.

[0004] NMN is widely found in natural foods, such as some fruits, vegetables, and mammalian milk. Literature reports that the NMN content in 100g of fresh avocado, cabbage, and broccoli samples is 6.519μg, 4.091μg, and 2.30μg, respectively. However, there is a problem with the low extraction rate of β-nicotinamide mononucleotide when extracting β-nicotinamide mononucleotide from natural foods. Summary of the Invention

[0005] The purpose of this disclosure is to provide a method for extracting β-nicotinamide mononucleotide from microalgae and a microalgae extract, which is simple in process and has a high extraction rate of β-nicotinamide mononucleotide.

[0006] To achieve the above objectives, the first aspect of this disclosure provides a method for extracting β-nicotinamide mononucleotide from microalgae, the method comprising: mixing microalgae raw material with an extractant for extraction treatment; wherein the microalgae raw material is selected from one or more of Chlorella, Spirulina and Haematococcus pluvialis; the extractant comprises 0.05-1.5% by weight of a nonionic surfactant, 0.5-1.0% by weight of an organic acid, 20-40% by weight of an alcohol and the balance being a solvent.

[0007] Optionally, the microalgae raw material includes Chlorella vulgaris, more preferably Chlorella proteoglycans; optionally, the microalgae raw material is used in the form of dried algae powder and fresh algae mud.

[0008] Optionally, the extractant comprises 0.5 to 1.5% by weight of a nonionic surfactant, 0.7 to 1.0% by weight of an organic acid, 30 to 40% by weight of an alcohol, and the balance being a solvent.

[0009] Optionally, the nonionic surfactant is selected from one or two of tea saponin and sitosterol; the organic acid is selected from one or more of citric acid, acetic acid and formic acid; the alcohol is selected from one or more of methanol and n-propanol; and the solvent is water.

[0010] Optionally, the extraction conditions include: the amount of the extractant used is 20-40 mL relative to 1 g of the microalgae raw material; the extraction temperature is 30-60 °C; the extraction time is 30-60 min; and optionally, the extraction is carried out under ultrasonic conditions with an ultrasonic power of 200-400 W.

[0011] Optionally, the extraction conditions include: the amount of the extractant is 30-40 mL relative to 1 g of the microalgae raw material; the extraction temperature is 40-60℃; the extraction time is 30-40 min; and optionally, the ultrasonic power is 300-400 W.

[0012] Optionally, the method further includes: centrifuging the product obtained from the extraction process, then taking the supernatant from the centrifugation process and ultrafiltration to obtain a filtrate; and freeze-drying the filtrate.

[0013] Optionally, the centrifugation conditions include: a centrifugation speed of 5000-10000 rpm, preferably 8000-10000 rpm, and a centrifugation time of 5-15 min, preferably 5-10 min; optionally, the ultrafiltration treatment uses an ultrafiltration membrane; the ultrafiltration membrane has a molecular weight cutoff of 1-5 kD, preferably 1-2 kD.

[0014] A second aspect of this disclosure provides a microalgae extract obtained according to the method of the first aspect of this disclosure, the microalgae extract comprising β-nicotinamide mononucleotide.

[0015] Optionally, the microalgae extract is a microalgae extract solution; the microalgae extract solution contains β-nicotinamide mononucleotide and the extractant; the content of β-nicotinamide mononucleotide is 2-200 mg / L based on the volume of the microalgae extract solution; preferably, when the microalgae raw material is Chlorella vulgaris, the content of β-nicotinamide mononucleotide is 100-200 mg / L based on the volume of the microalgae extract solution.

[0016] Through the above technical solution, this disclosure provides a method for extracting β-nicotinamide mononucleotide from microalgae and a microalgae extract, wherein the microalgae raw material is easy to cultivate, grows rapidly, has high protein content, and has a high β-nicotinamide mononucleotide content; the extraction method of this disclosure is simple, has a high extraction rate, and is low in cost.

[0017] Other features and advantages of this disclosure will be described in detail in the following detailed description section. Attached Figure Description

[0018] The accompanying drawings are provided to further illustrate the present disclosure and form part of the specification. They are used together with the following detailed description to explain the present disclosure, but do not constitute a limitation thereof. In the drawings:

[0019] Figure 1 This is a chromatogram of β-nicotinamide mononucleotide in the microalgae extract obtained in Example 1.

[0020] Figure 2 This is a standard curve of NMN mass concentration versus peak area. Detailed Implementation

[0021] The specific embodiments of this disclosure will be described in detail below with reference to the accompanying drawings. It should be understood that the specific embodiments described herein are for illustration and explanation only and are not intended to limit this disclosure.

[0022] The inventors of this disclosure discovered through experiments that different types of microalgae have high β-nicotinamide mononucleotide (NMN) content, which is much higher than that of known fruits and vegetables used for NMN extraction, making them suitable as raw materials for NMN extraction.

[0023] The first aspect of this disclosure provides a method for extracting β-nicotinamide mononucleotide from microalgae, the method comprising: mixing microalgae raw material with an extractant for extraction treatment;

[0024] The microalgae raw material is selected from one or more of Chlorella, Spirulina and Haematococcus pluvialis; the extractant includes 0.05-1.5% by weight of nonionic surfactant, 0.5-1.0% by weight of organic acid, 20-40% by weight of alcohol and the balance being solvent.

[0025] This disclosure provides a method for extracting β-nicotinamide mononucleotide from microalgae. The microalgae raw material is easy to cultivate, grows rapidly, has high protein content, and also has a high β-nicotinamide mononucleotide content. The extraction method disclosed herein is simple, has a high extraction rate, and is low in cost.

[0026] In this disclosure, the content of β-nicotinamide mononucleotide was detected by high-performance liquid chromatography-mass spectrometry. Specific instrument conditions can be adopted as disclosed in the literature "Simultaneous quantification of nicotinamide mononucleotide and related pyridine compounds in mouse tissues by UHPLC–MS / MS".

[0027] In a preferred embodiment, the microalgae raw material is selected from Chlorella vulgaris, and more preferably from Chlorella proteoglycans. This disclosure utilizes Chlorella proteoglycans with a higher β-nicotinamide mononucleotide content for extraction to obtain an extract with a higher NMN content.

[0028] In one embodiment, the microalgae raw material is used in the form of dried algae powder and fresh algae mud.

[0029] The microalgae raw materials (such as algal strains, dried algal powder, etc.) used in this disclosure can be microalgae of known species in the art that can be obtained through conventional channels, and can be propagated and cultured using conventional methods. Alternatively, the microalgae raw materials can be pretreated using methods known in the art before extraction and processing.

[0030] In one specific embodiment, the Chlorella strain expansion treatment adopts the method disclosed in the literature "Wang Q, Yu Z, Wei D, et al. Mixotrophic Chlorella pyrenoidosa as cell factory for ultrahigh-efficient removal of ammonium from catalyzer wastewater with valuable algal biomassco production through short-time acclimation[J]. Bioresource Technology, 2021, 333: 125151"; the Spirulina strain expansion treatment adopts the method disclosed in the literature "Li Xufeng, Wang Changhai, Wen Shaohong. Study on Spirulina culture conditions[J]. Food and Fermentation Industries, 1999, 25(4): 13".

[0031] In one specific embodiment, the microalgae raw material is in the form of dried algae powder, which is obtained by expanding, harvesting and freeze-drying the algae strain; wherein the parameters in the process of preparing the dried algae powder are conventional process parameters in the art.

[0032] In a preferred embodiment, the extractant comprises 0.5–1.5 wt% of a nonionic surfactant, 0.7–1.0 wt% of an organic acid, 30–40 wt% of an alcohol, and the balance being a solvent. Using an extractant with the preferred composition can achieve a higher NMN extraction rate.

[0033] In one specific embodiment, the nonionic surfactant is selected from one or both of tea saponin and sitosterol; more preferably, it is tea saponin.

[0034] The organic acid is selected from one or more of citric acid, acetic acid and formic acid;

[0035] The alcohol is selected from one or more of methanol and n-propanol;

[0036] The solvent is water.

[0037] The extraction agent provided in this embodiment can further improve the NMN extraction rate.

[0038] In one embodiment, the extraction conditions include: the amount of the extractant used is 20-40 mL relative to 1 g of the microalgae raw material; the extraction temperature is 30-60 °C; and the extraction time is 30-60 min.

[0039] In one specific embodiment, the extraction process is performed under ultrasonic conditions, with an ultrasonic power of 200–400 W. In this disclosure, "ultrasonic conditions" can refer to ultrasonic instruments well-known in the art.

[0040] In a preferred embodiment, the extraction conditions include: the amount of the extractant is 30-40 mL relative to 1 g of the microalgae raw material; the extraction temperature is 40-60°C; the extraction time is 30-40 min; and optionally, the ultrasonic power is 300-400 W. The preferred extraction conditions provided in this disclosure are more suitable for extracting β-nicotinamide mononucleotide from microalgae.

[0041] In one embodiment, the method further includes: centrifuging the product obtained from the extraction process, and then taking the supernatant from the centrifugation process for ultrafiltration to obtain a filtrate;

[0042] The filtrate is then freeze-dried.

[0043] In one specific embodiment, the centrifugation conditions include: a centrifugation speed of 5000-10000 rpm, preferably 8000-10000 rpm, and a centrifugation time of 5-15 min, preferably 5-10 min.

[0044] In one specific embodiment, the ultrafiltration treatment employs an ultrafiltration membrane; the ultrafiltration membrane has a molecular weight cutoff of 1–5 kD, preferably 1–2 kD.

[0045] In one embodiment, the β-nicotinamide mononucleotide extraction rate of the method provided in this disclosure is 0.005–0.5% by weight; preferably 0.35–0.5% by weight.

[0046] A second aspect of this disclosure provides a microalgae extract obtained according to the method described in the first aspect of this disclosure, the microalgae extract comprising β-nicotinamide mononucleotide.

[0047] In one embodiment, the microalgae extract is a microalgae extract solution; the microalgae extract solution contains β-nicotinamide mononucleotide and the extractant; the β-nicotinamide mononucleotide content is 2-200 mg / L based on the volume of the microalgae extract solution.

[0048] In a preferred embodiment, when the microalgae raw material is Chlorella, the content of β-nicotinamide mononucleotide is 100-200 mg / L based on the volume of the microalgae extract.

[0049] The following detailed embodiments further illustrate the above-described content of this disclosure. However, this should not be construed as limiting the scope of this disclosure to the following embodiments.

[0050] The raw material sources used in the following examples and comparative examples include:

[0051] Chlorella powder with protein nucleus is obtained through the following steps:

[0052] The *Chlorella pyrenoidosa* strain (purchased from the algal strain bank of the Research Institute of Petroleum Processing, Sinopec) was used for propagation treatment. The propagation treatment followed the method disclosed in the literature "Wang Q, Yu Z, Wei D, et al. Mixotrophic *Chlorella pyrenoidosa* as cell factory for ultrahigh-efficient removal of ammonium from catalyst wastewater with valuable algal biomass coproduction through short-time acclimation[J]. Bioresource Technology, 2021, 333:125151". The culture medium used for propagation was Basal medium containing 10 g / L glucose. The culture conditions were: light intensity 100 μmol / (m²). 2Under continuous white LED illumination (·s), the algae were cultured for 5 days in a shaking incubator at 30℃ and 150r / min. Then, the expanded algae were centrifuged (5000rmp, 5min) and harvested, and then freeze-dried (FD-1C-80 Beijing Bilang Experimental Equipment Co., Ltd.) to produce dry algae powder. The freeze-drying conditions included: cold trap temperature of -80℃; vacuum degree of less than 3Pa; and drying time of 48h.

[0053] Spirulina powder is obtained through the following steps:

[0054] Spirulina strains (purchased from the strain bank of the Research Institute of Petroleum Processing, China Petrochemical Corporation) were used for propagation treatment; the propagation treatment followed the method disclosed in the literature "Li Xufeng, Wang Changhai, Wen Shaohong. Study on culture conditions of Spirulina [J]. Food and Fermentation Industries, 1999, 25(4):13"; the culture medium used for propagation was Zarrouk medium; the culture conditions were: light intensity of 50 μmol / (m 2 Spirulina was cultured for 20 days in a shaking incubator at 25°C and 150 r / min under continuous white LED illumination. Then, the expanded culture Spirulina was harvested and freeze-dried under the same harvesting and freeze-drying conditions as Chlorella pyrenoidosa powder.

[0055] The Haematococcus pluvialis powder was purchased from Yunnan Shiprui Biotechnology Co., Ltd.

[0056] Tea saponin, citric acid, and Tween-20 were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0057] The ultrafiltration membrane was purchased from Shandong Bona Biotechnology Group Co., Ltd., with a molecular weight cutoff of 1 kDa.

[0058] Example 1

[0059] (1) Take 1g of Chlorella pyrenoidosa powder and mix it evenly with the extraction solvent, and sonicate at 300W for 50min at 50℃.

[0060] (2) Centrifuge the mixture extracted in step (1), and ultrafilter the supernatant obtained after centrifugation, with a molecular weight cutoff of 1KD, and collect the filtrate;

[0061] (3) The filtrate from step (2) is freeze-dried under vacuum to obtain the final product.

[0062] The amount of extractant used is 30 mL relative to 1 g of Chlorella protein powder.

[0063] The extractant contains: 1.5% by weight tea saponin, 0.8% by weight citric acid, 20% by weight methanol, and the balance is water.

[0064] The chromatogram of the extracted product obtained in Example 1 is shown below. Figure 1As shown in the figure, a peak eluted at 1.16 min, which matches the molecular weight of β-nicotinamide mononucleotide (NMN) 334.2192, indicating that the extract obtained by the method provided in this disclosure contains NMN.

[0065] Example 2

[0066] (1) Take 1g of Chlorella pyrenoidosa powder and mix it evenly with the extraction solvent, and sonicate at 400W for 40min at 60℃.

[0067] (2) Centrifuge the mixture extracted in step (1), and ultrafilter the supernatant obtained after centrifugation, with a molecular weight cutoff of 1KD, and collect the filtrate;

[0068] (3) The filtrate from step (2) is freeze-dried under vacuum to obtain the final product.

[0069] The amount of extractant used is 25 ml, which is equivalent to 1 g of Chlorella protein powder.

[0070] The extractant contains: 1.0 wt% tea saponin, 1.0 wt% citric acid, 30 wt% methanol, and the balance is water.

[0071] Example 3

[0072] (1) Take 1g of Chlorella pyrenoidosa powder and mix it evenly with the extraction solvent, and sonicate at 200W for 60min at 40℃.

[0073] (2) Centrifuge the mixture extracted in step (1), and ultrafilter the supernatant obtained after centrifugation, with a molecular weight cutoff of 1KD, and collect the filtrate;

[0074] (3) The filtrate from step (2) is freeze-dried under vacuum to obtain the final product.

[0075] The amount of extractant used is 40 ml, which is equivalent to 1 g of Chlorella protein powder.

[0076] The extractant contains: 0.8% by weight tea saponin, 0.7% by weight citric acid, 40% by weight methanol, and the balance is water.

[0077] Example 4

[0078] (1) Take 1g of Chlorella pyrenoidosa powder and mix it evenly with the extraction solvent, and sonicate at 350W for 50min at 50℃.

[0079] (2) Centrifuge the mixture extracted in step (1), and ultrafilter the supernatant obtained after centrifugation to a molecular weight cutoff of 1.5KD, and collect the filtrate;

[0080] (3) The filtrate from step (2) is freeze-dried under vacuum to obtain the final product.

[0081] The amount of extractant used is 20 ml, which is equivalent to 1 g of Chlorella protein powder.

[0082] The extractant contains: 0.5% by weight tea saponin, 1.0% by weight citric acid, 30% by weight methanol, and the balance is water.

[0083] Example 5

[0084] The extraction method used is the same as in Example 1, except that:

[0085] Use Tween-20 instead of tea saponin, with the dosage remaining the same.

[0086] Example 6

[0087] The extraction method used is the same as in Example 1, except that:

[0088] The amount of extractant used is 10 ml, which is equivalent to 1 g of Chlorella protein powder.

[0089] Example 7

[0090] The extraction method used is the same as in Example 1, except that:

[0091] The microalgae raw material is spirulina powder.

[0092] Example 8

[0093] The extraction method used is the same as in Example 1, except that:

[0094] The microalgae raw material is Haematococcus pluvialis powder.

[0095] Example 9

[0096] The extraction method used is the same as in Example 1, except that:

[0097] The extractant was replaced with a mixture comprising CTAB, lauramide propyl hydroxysulfobetaine, and formic acid, wherein the mixture contained 1.5% by weight of hexadecyltrimethylamine bromide (CTAB), 0.3% by weight of sulfobetaine, and 0.5% by weight of formic acid, with the remainder being water.

[0098] Example 10

[0099] The extraction method used is the same as in Example 2, except that:

[0100] The extractant contains: 0.3% by weight tea saponin, 0.5% by weight citric acid, 25% by weight methanol, and the balance is water.

[0101] Example 11

[0102] The extraction method used is the same as in Example 2, except that:

[0103] The amount of extractant used is 35ml relative to 1g of Chlorella protein powder.

[0104] Example 12

[0105] The extraction method used is the same as in Example 11, except that:

[0106] The extraction conditions included an extraction temperature of 35℃, an extraction time of 60 min, and an ultrasonic power of 200W.

[0107] Example 13

[0108] The extraction method used is the same as in Example 1, except that:

[0109] The extractant contains: 1.5% by weight sitosterol, 0.8% by weight acetic acid, 20% by weight n-propanol, and the balance is water.

[0110] Comparative Example 1

[0111] The extraction method used was the same as in Example 1, except that the extract did not contain tea saponins and citric acid, and only an aqueous methanol solution with a methanol concentration of 20% by weight was used.

[0112] Comparative Example 2

[0113] The same extraction method as in Example 1 was used, except that the extraction solvent contained 5% by weight tea saponin, 3% by weight citric acid, 30% by weight methanol, and the remainder was water.

[0114] Comparative Example 3

[0115] A method for extracting NMN from fruits and vegetables includes the following steps:

[0116] (1) Crush the edamame;

[0117] (2) Add the powder obtained after pulverization to ammonia water, stir and mix evenly, then add a mixed solution of ethylenediaminetetraacetic acid (EDTA) and sodium chloride, and sonicate at 200W for 10 minutes; wherein, the ratio of powder to mixed solution is 1:5; the EDTA content in the mixed solution is 1% by weight, and the sodium chloride content is 1.5% by weight.

[0118] (3) After the ultrasound in step (2) is completed, continue to add a mixture including CTAB, lauramide propyl hydroxysulfonate betaine and formic acid, and sonicate at 800W and 60℃ for 80 minutes; wherein, the ratio of powder to mixture is 1:22; the content of CTAB is 1.5% by weight; the content of sulfonate betaine is 0.3% by weight; and the content of formic acid is 0.5% by weight.

[0119] (4) Filter the product obtained in step (3) and collect the supernatant, concentrate and crystallize and dry the solid product.

[0120] The microalgae extracts obtained in the above examples and comparative examples were subjected to high-performance liquid chromatography-mass spectrometry (HPLC-MS / MS) (Agilent 1260 Infinity) to determine the β-nicotinamide mononucleotide content. The results are listed in Table 1 below. A standard curve of NMN mass concentration versus peak area was established (e.g., ...). Figure 2 The NMN content in the microalgae extract obtained from each 1g of microalgae raw material (the mass of the microalgae raw material is expressed as DW on a dry weight basis) is determined (the data corresponding to the column of β-nicotinamide mononucleotide (mg / g DW) in Table 1 below), and then the NMN extraction rate of the microalgae raw material is obtained. This disclosure uses the chromatography-mass spectrometry test conditions disclosed in the literature "Simultaneous quantification of nicotinamide mononucleotide and related pyridine compounds in mouse tissues by UHPLC–MS / MS".

[0121] Table 1

[0122]

[0123]

[0124] The microalgae extract is in the form of an extract, containing β-nicotinamide mononucleotide and the extraction agent used. The mass of NMN extracted (relative to 1g of algal powder raw material) is detected by liquid chromatography-mass spectrometry. The extraction rate = mass of extracted NMN / mass of algal powder. The NMN content in the extract = mass of extracted NMN / volume of extract. In this embodiment, the mass (dry weight) of algal powder is 1g.

[0125] Based on the data in Table 1 above: Comparing Example 1 with Comparative Examples 1 and 2, the composition of the extractant used in Example 1 meets the requirements of "0.05-1.5% by weight of nonionic surfactant, 0.5-1.0% by weight of organic acid, 20-40% by weight of alcohol and the balance solvent". Example 1 has a higher β-nicotinamide mononucleotide extraction rate and β-nicotinamide mononucleotide content.

[0126] Comparing Examples 1-9 with Comparative Example 3, the method provided in this disclosure can achieve a higher β-nicotinamide mononucleotide extraction rate compared to existing methods for extracting NMN from fruits and vegetables.

[0127] Comparing Example 1 with Example 5, Example 1 uses tea saponin and has a higher β-nicotinamide mononucleotide extraction rate and β-nicotinamide mononucleotide content;

[0128] Comparing Example 1 with Example 6, in Example 1, the amount of extractant used relative to 1g of microalgae raw material was 20-40mL, and Example 1 had a higher β-nicotinamide mononucleotide extraction rate;

[0129] Comparing Example 1 with Examples 7 and 8, Example 1 uses Chlorella powder for extraction and has a higher β-nicotinamide mononucleotide extraction rate and β-nicotinamide mononucleotide content.

[0130] Example 1 was compared with Example 9. The extractant in Example 1 included tea saponin, citric acid, methanol and water. Example 1 had a higher β-nicotinamide mononucleotide extraction rate and β-nicotinamide mononucleotide content.

[0131] Comparing Example 1 with Example 10, the extractant composition of Example 10 satisfies "including 0.5-1.5% by weight of nonionic surfactant, 0.7-1.0% by weight of organic acid, 30-40% by weight of alcohol and the balance solvent". Example 10 has a higher β-nicotinamide mononucleotide extraction rate and β-nicotinamide mononucleotide content.

[0132] Comparing Example 2 with Example 11, in Example 2, the amount of extractant used was 25 ml relative to 1 g of Chlorella proteinensis powder, and Example 2 had a higher β-nicotinamide mononucleotide extraction rate;

[0133] Further comparing Example 11 with Example 12, the extraction conditions in Example 11 met the requirements of "extraction temperature of 40-60℃; extraction time of 30-40 min; ultrasonic power of 300-400 W". Example 11 had a higher β-nicotinamide mononucleotide extraction rate and β-nicotinamide mononucleotide content.

[0134] The preferred embodiments of this disclosure have been described in detail above with reference to the accompanying drawings. However, this disclosure is not limited to the specific details of the above embodiments. Within the scope of the technical concept of this disclosure, various simple modifications can be made to the technical solutions of this disclosure, and these simple modifications all fall within the protection scope of this disclosure.

[0135] It should also be noted that the various specific technical features described in the above embodiments can be combined in any suitable manner without contradiction. To avoid unnecessary repetition, this disclosure will not describe the various possible combinations separately.

[0136] Furthermore, various different embodiments of this disclosure can be combined in any way, as long as they do not violate the spirit of this disclosure, they should also be regarded as the content disclosed in this disclosure.

Claims

1. A method for extracting β-nicotinamide mononucleotide from microalgae, characterized in that, This method includes: mixing microalgae raw materials with an extractant for extraction; The microalgae raw material is selected from one or more of Chlorella, Spirulina and Haematococcus pluvialis; The extractant comprises 0.05-1.5% by weight of a nonionic surfactant, 0.5-1.0% by weight of an organic acid, 20-40% by weight of an alcohol, and the balance being a solvent. The nonionic surfactant is selected from one or two of tea saponin and sitosterol; the organic acid is selected from one or more of citric acid, acetic acid and formic acid; the alcohol is selected from one or more of methanol and n-propanol; and the solvent is water.

2. The method according to claim 1, characterized in that, The microalgae raw material is selected from Chlorella.

3. The method according to claim 2, characterized in that, The microalgae raw material is Chlorella proteoglycans.

4. The method according to claim 1, characterized in that, The microalgae raw materials are used in the form of dried algae powder and fresh algae mud.

5. The method according to claim 1, characterized in that, The extractant comprises 0.5-1.5% by weight of a nonionic surfactant, 0.7-1.0% by weight of an organic acid, 30-40% by weight of an alcohol, and the balance being a solvent.

6. The method according to claim 1, characterized in that, The extraction conditions include: the amount of the extractant used is 20-40 mL relative to 1 g of the microalgae raw material; the extraction temperature is 30-60℃; and the extraction time is 30-60 min.

7. The method according to claim 6, characterized in that, The extraction process is performed under ultrasonic conditions with an ultrasonic power of 200-400W.

8. The method according to claim 6, characterized in that, The extraction conditions include: the amount of the extractant used is 30-40 mL relative to 1 g of the microalgae raw material; the extraction temperature is 40-60℃; and the extraction time is 30-40 min.

9. The method according to claim 7, characterized in that, The extraction and processing conditions include: ultrasonic power of 300~400W.

10. The method according to claim 1, characterized in that, The method further includes: centrifuging the product obtained from the extraction process, and then taking the supernatant from the centrifugation process for ultrafiltration to obtain the filtrate; The filtrate is then freeze-dried.

11. The method according to claim 10, characterized in that, The centrifugation conditions include: a centrifugation speed of 5000~10000 rpm and a centrifugation time of 5~15 min.

12. The method according to claim 11, characterized in that, The centrifugation conditions include: a centrifugation speed of 8000~10000 rpm and a centrifugation time of 5~10 min.

13. The method according to claim 10, characterized in that, The ultrafiltration process uses an ultrafiltration membrane; the molecular weight cutoff of the ultrafiltration membrane is 1~5kD.

14. The method according to claim 13, characterized in that, The ultrafiltration membrane has a molecular weight cutoff of 1~2 kD.

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