Saccharomyces cerevisiae with improved efficiency of xylose utilization and its application and method for producing ethanol

Abstract

The present application relates to the technical field of bioengineering, and discloses a Saccharomyces cerevisiae with improved utilization efficiency of xylose, application of the Saccharomyces cerevisiae and a method for producing ethanol. The Saccharomyces cerevisiae provided by the present application has improved growth ability and utilization efficiency of xylose in the process of glucose-xylose co-fermentation, so that the production cost is effectively reduced when the Saccharomyces cerevisiae is used to produce ethanol by using xylose-containing substrates such as lignocellulose hydrolysate, and the production efficiency is improved, so that the Saccharomyces cerevisiae has great production and application value.

Description

Technical Field

[0001] This invention relates to the field of bioengineering technology, specifically to a brewing yeast that improves xylose utilization efficiency, its application, and a method for producing ethanol. Background Technology

[0002] Petrochemical resources are non-renewable, and socio-economic development based on them faces risks. Furthermore, their extraction and absolute carbon emissions place immense pressure on the environment. Using biomass as a raw material to produce fuels and chemicals is an inevitable trend. Whether it's the current production method primarily using low-cost food crops like corn as raw materials, or the production method promoted by various countries using lignocellulosic raw materials such as agricultural and forestry waste, both methods feature renewable raw materials, less pollution during production, and a carbon cycle due to carbon sequestration during production and carbon release during combustion, resulting in lower overall carbon emissions.

[0003] Currently, the main process for producing fuels and chemicals from biomass involves converting starch from low-priced grain crops like corn into glucose, which is then fermented to produce fuel ethanol and other products. However, this model competes with humans and livestock for food, and production costs are gradually increasing as grain prices rise. In contrast, production routes using lignocellulosic raw materials such as agricultural and forestry waste can fully utilize resources, promote employment, and optimize the industrial structure, thus attracting widespread attention.

[0004] Lignocellulose raw materials are mainly composed of cellulose and hemicellulose, which, after hydrolysis, yield glucose and xylose in a ratio of approximately 2:1. Full utilization of these two sugars is fundamental to improving raw material utilization and reducing production costs. However, the safe and efficient glucose fermentation strain, *Saccharomyces cerevisiae*, cannot metabolize xylose. Through metabolic pathway engineering, the xylose metabolic pathway can be introduced and optimized, enabling *Saccharomyces cerevisiae* to utilize xylose. However, xylose utilization efficiency is far lower than glucose utilization efficiency, and the presence of glucose has a significant negative impact on xylose utilization. Therefore, improving the efficiency of xylose utilization by recombinant strains in glucose-xylose mixed fermentation has always been a technical problem that needs to be solved in this field. Summary of the Invention

[0005] The purpose of this invention is to overcome the aforementioned problems in the prior art and provide a brewing yeast with improved xylose utilization efficiency, its application, and a method for producing ethanol. The brewing yeast provided by this invention improves xylose utilization efficiency during glucose-xylose co-fermentation and also has the advantage of increasing ethanol accumulation.

[0006] To achieve the above objectives, the present invention provides a brewing yeast with improved xylose utilization efficiency, wherein the brewing yeast is selected from brewing yeasts that have a xylose metabolic pathway and whose transcription factor Skn7 activity is inhibited.

[0007] A second aspect of the present invention provides a method for improving the growth ability of Saccharomyces cerevisiae to utilize xylose and / or improving the utilization efficiency of xylose by Saccharomyces cerevisiae, the method comprising inhibiting the activity of transcription factor Skn7 in Saccharomyces cerevisiae, wherein the Saccharomyces cerevisiae is selected from Saccharomyces cerevisiae with a xylose metabolic pathway.

[0008] The third aspect of the present invention provides the application of the brewing yeast as described in the first aspect above and / or the method as described in the second aspect above in ethanol production, particularly in the application of ethanol fermentation production using a substrate containing lignocellulose hydrolysate.

[0009] A fourth aspect of the present invention provides a method for producing ethanol, the method comprising fermenting a xylose-containing substrate with Saccharomyces cerevisiae and inhibiting the activity of transcription factor Skn7 in the Saccharomyces cerevisiae, wherein the Saccharomyces cerevisiae is selected from Saccharomyces cerevisiae with a xylose metabolic pathway.

[0010] Through the above technical solution, the present invention can achieve the following beneficial effects:

[0011] (1) When the brewing yeast provided by the present invention is fermented using a substrate containing xylose and glucose and other sugars, the growth of the strain and the efficiency of xylose utilization are effectively improved. This solves the problem of low xylose utilization efficiency of existing xylose-utilizing strains in glucose-xylose co-fermentation system, while also improving the growth rate of the strain, thereby accelerating the fermentation process and improving production efficiency.

[0012] (2) The brewing yeast provided by the present invention can also increase the accumulation of ethanol during glucose xylose co-fermentation, thereby improving the ethanol yield and production efficiency when using the brewing yeast for fermentation production, thus improving the raw material utilization rate and product yield, and reducing production costs. It is very suitable for large-scale industrial production applications. Attached Figure Description

[0013] Figure 1 This is a schematic diagram of the structure of the DNA fragment dSKN7-KanMX used for gene knockout in Example 1.

[0014] Figure 2 This is a comparison diagram of the effects of knocking out the SKN7 gene on the ethanol fermentation of Saccharomyces cerevisiae BSGX001 with different carbon sources in Example 3. Detailed Implementation

[0015] The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0016] In this invention, recombinant yeast BSGX001, or Saccharomyces cerevisiae BSGX001, refers to the Saccharomyces cerevisiae with accession number CGMCC No. 17264, which has been disclosed in CN201910304995.3. Saccharomyces cerevisiae SDIC Sc E01 refers to the Saccharomyces cerevisiae with accession number GDMCC No: 62491, which has been disclosed in CN202210813571.1.

[0017] In this invention, transcription factor Skn7 refers to a transcription factor that responds to heat shock and oxidative stress, and the nucleotide sequence of its encoding gene (SKN7 gene) is accessed in GenBank with the accession number Gene ID: 856613.

[0018] In this invention, inhibiting the activity of transcription factor Skn7 means knocking out or mutating the SKN7 gene, or interfering with the expression of the SKN7 gene, so that the transcription factor cannot be expressed normally, or the activity of the expressed transcription factor Skn7 decreases or is lost ("decreases or is lost" means that the activity of transcription factor Skn7 is reduced to 20% or less of the original activity, for example, reduced to 20%, 18%, 15%, 12%, 10%, 5%, 2%, 1%, 0.5%, 0.1%, 0, or any intermediate value of any two of the above values).

[0019] The inventors of this invention discovered that while introducing a xylose metabolic pathway into *Saccharomyces cerevisiae* enables the recombinant *Saccharomyces cerevisiae* to utilize xylose, thus improving the production efficiency and yield of ethanol fermentation using xylose-containing substrates such as lignocellulose hydrolysate, the presence of other sugars in the substrate (such as glucose, mannose, and galactose) negatively impacts xylose metabolism. Moreover, this negative impact persists even after the metabolism of these sugars is exhausted. Therefore, improving the utilization efficiency of xylose-containing substrates by recombinant yeast is essential to further enhance xylose utilization. Through research, the inventors ingeniously discovered that inhibiting the activity of transcription factor Skn7 in recombinant yeast with the introduced xylose metabolic pathway effectively improves the utilization efficiency of xylose in substrates containing both xylose and other sugars such as glucose. Further research revealed that inhibiting the activity of transcription factor Skn7 not only improves the utilization efficiency of xylose in recombinant brewing with xylose metabolic pathways in xylose co-fermentation systems with other sugars, but also further improves the growth of microorganisms and increases ethanol accumulation.

[0020] Based on the above findings, the first aspect of the present invention provides a brewing yeast with improved xylose utilization efficiency, wherein the brewing yeast is selected from brewing yeasts that have a xylose metabolic pathway and whose transcription factor Skn7 activity is inhibited.

[0021] According to a preferred embodiment of the present invention, the brewer's yeast is selected from brewer's yeast CGMCC No. 17264 and / or GDMCC No: 62491 (with inhibited transcription factor Skn7 activity).

[0022] In this invention, there are no particular limitations on the specific method for inhibiting the transcription factor Skn7 in Saccharomyces cerevisiae; any existing method in the art capable of achieving this objective is applicable. Examples include knocking out the SKN7 gene, mutating the SKN7 gene to inactivate its expressed protein, or introducing other exogenous genes to interfere with the expression of the SKN7 gene.

[0023] A second aspect of the present invention provides a method for improving the growth ability of Saccharomyces cerevisiae to utilize xylose and / or improving the utilization efficiency of xylose by Saccharomyces cerevisiae, the method comprising inhibiting the activity of transcription factor Skn7 in Saccharomyces cerevisiae, wherein the Saccharomyces cerevisiae is selected from Saccharomyces cerevisiae with a xylose metabolic pathway.

[0024] In this invention, there are no particular limitations on the method of inhibiting the activity of transcription factor Skn7 in Saccharomyces cerevisiae, as long as its activity is reduced or lost (to the aforementioned degree). This can be achieved by modifying Saccharomyces cerevisiae to inhibit the activity of transcription factor Skn7, or by adding additional reagents during the production process to inhibit the activity of transcription factor Skn7.

[0025] According to a preferred embodiment of the present invention, the methods for inhibiting the activity of transcription factor Skn7 in *Saccharomyces cerevisiae* include: knocking out the SKN7 gene, mutating the SKN7 gene to render the expressed protein inactive, or interfering with the expression of the SKN7 gene, thereby preventing the expression of transcription factor Skn7 or reducing or eliminating the activity of the expressed transcription factor Skn7. In this invention, methods for interfering with the expression of the SKN7 gene may include adding substances that interfere with the expression of the SKN7 gene during fermentation, or further modifying *Saccharomyces cerevisiae* to introduce other exogenous genes that interfere with the expression of the SKN7 gene.

[0026] According to a preferred embodiment of the present invention, the activity of transcription factor Skn7 in Saccharomyces cerevisiae is inhibited by knocking out the SKN7 gene.

[0027] According to a preferred embodiment of the present invention, the Saccharomyces cerevisiae with the xylose metabolic pathway is selected from Saccharomyces cerevisiae CGMCC No. 17264 and / or GDMCC No: 62491.

[0028] A third aspect of this invention provides the application of the brewing yeast described in the first aspect and / or the method described in the second aspect above in ethanol production. In particular, it is applied in ethanol fermentation using substrates containing lignocellulose hydrolysates. For example, brewing yeast with the above characteristics can be used to ferment substrates containing xylose and other sugars such as glucose as carbon sources for ethanol production. Alternatively, brewing yeast can be modified using the methods described above, and then the modified brewing yeast can be used for ethanol fermentation. Or, when using brewing yeast with a xylose metabolic pathway to ferment substrates containing xylose and other sugars such as glucose as carbon sources for ethanol production, a substance that interferes with the expression of the SKN7 gene can be added to improve the efficiency of xylose utilization and the growth rate of the strain during the production process.

[0029] A fourth aspect of the present invention provides a method for producing ethanol, the method comprising fermenting a xylose-containing substrate with Saccharomyces cerevisiae and inhibiting the activity of transcription factor Skn7 in the Saccharomyces cerevisiae, wherein the Saccharomyces cerevisiae is selected from Saccharomyces cerevisiae with a xylose metabolic pathway.

[0030] In this invention, there are no particular limitations on the source and specific composition of the substrate, as long as the brewing yeast can utilize it for ethanol fermentation. According to a preferred embodiment of the invention, the substrate comprises a lignocellulose hydrolysate. For example, the lignocellulose hydrolysate can be used directly as the fermentation substrate, or other reagents can be added to the lignocellulose hydrolysate to prepare a substrate for brewing yeast to carry out ethanol fermentation.

[0031] When lignocellulose hydrolysate is used as a fermentation substrate (one of the components), the sugars such as glucose, mannose, and galactose contained in the hydrolysate can be utilized by Saccharomyces cerevisiae (through its natural utilization pathway) and converted into ethanol. However, the presence of these sugars negatively impacts xylose utilization, thus affecting the overall ethanol production efficiency and yield. Inhibiting the activity of transcription factor Skn7 improves the utilization efficiency of xylose by Saccharomyces cerevisiae and the growth rate of the strain, thereby increasing the raw material utilization rate and production efficiency compared to when Skn7 activity is not inhibited. Simultaneously, it has no adverse effect on the utilization efficiency of other sugars available to Saccharomyces cerevisiae. Therefore, preferably, the substrate may also contain at least one of glucose, mannose, and galactose as a carbon source.

[0032] According to a preferred embodiment of the present invention, the weight ratio of glucose to xylose in the substrate is 1:0.5-5.

[0033] Preferably, the weight ratio of glucose to xylose is 1:0.5-1.5. For example, it can be 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, or any intermediate value between any two of the above ratios.

[0034] According to a preferred embodiment of the present invention, the activity of transcription factor Skn7 in the *Saccharomyces cerevisiae* is inhibited by knocking out the SKN7 gene, mutating the SKN7 gene to inactivate the protein it expresses, or interfering with the expression of the SKN7 gene. Knocking out the SKN7 gene is preferred.

[0035] According to a preferred embodiment of the present invention, the Saccharomyces cerevisiae with the xylose metabolic pathway is selected from Saccharomyces cerevisiae CGMCC No. 17264 and / or GDMCC No: 62491.

[0036] In this invention, there are no particular limitations on the specific fermentation methods and conditions. To further improve ethanol production efficiency, according to a preferred embodiment of the invention, the fermentation conditions include an initial OD... 600=0.5-5, rotation speed 100-300 rpm, temperature 25-35℃. The preferred fermentation time is 24-72 hours.

[0037] Preferably, the fermentation conditions include initial OD 600 =0.5-2, speed 150-250rpm, temperature 27-32℃.

[0038] The present invention will be described in detail below through embodiments. It should be understood that the following embodiments are only used to further explain and illustrate the content of the present invention by way of example, and are not intended to limit the present invention.

[0039] In the following examples, the starting strain BSGX001 (abbreviated as BSGX001) is a recombinant Saccharomyces cerevisiae strain with an introduced and optimized xylose metabolic pathway, its accession number is CGMCC No. 17264, and the specific construction method is described in CN201910304995.3. The starting strain SDIC Sc E01 (abbreviated as DIC Sc E01) is a recombinant Saccharomyces cerevisiae strain that can tolerate relatively harsh fermentation environments and can utilize xylose and glucose for ethanol fermentation production more quickly, its accession number is GDMCC No:62491, and the specific construction method is described in CN202210813571.1. Unless otherwise specified, all reagents or materials used were commercially available products purchased from reputable biological or chemical reagent / material suppliers, and all reagents were of analytical grade.

[0040] The culture media used in the following examples are as follows:

[0041] YPD medium: Weigh the raw materials according to the ratio of 2% peptone, 1% yeast extract, and 2% glucose, and dissolve them in deionized water. Autoclave at 115℃ for 30 minutes, then cool to obtain YPD liquid medium. Add 2% agar powder before sterilization. After sterilization, wait until it is cool to a comfortable temperature (about 50℃), then pour into plates. Allow to cool and solidify to obtain YPD culture plates.

[0042] Glucose-xylose glycosaccharide fermentation medium (using glucose and xylose as carbon sources): Weigh the raw materials according to the following ratio: yeast basic nitrogen source (YNB) 1.7 g / L, (NH4)2SO4 5 g / L, CSM-URA 0.77 g / L, and dissolve them in deionized water to prepare Solution 1. Simultaneously, prepare 40% by weight glucose solution and 40% by weight xylose solution. Autoclave each solution at 115℃ for 30 min. After cooling, add the above sugar solutions to Solution 1 to a final concentration of 20 g / L glucose and 20 g / L xylose to obtain the glucose-xylose glycosaccharide fermentation liquid medium.

[0043] In the following examples, conventional techniques and methods used in the fields of genetic engineering and molecular biology were employed to modify the starting strain BSGX001, thereby inhibiting the activity of its transcription factor Skn7 by knocking out the SKN7 gene. Specific methods can be found in Methods in yeast genetics and genomics: a Cold Spring Harbor Laboratory course manual 2015 edition (Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press, 2005).

[0044] Example 1: PCR to obtain DNA fragments for gene knockout

[0045] Using plasmid pUG6 (GenBank: AF298793.1) as a template, and dSKN7s and dSKN7a as primers (see Table 1 for specific sequences), the KanMX expression cassette DNA fragment dSKN7-KanMX containing the SKN7 homologous recombination arm was amplified using the system and conditions in Table 1 below.

[0046] Table 1

[0047]

[0048] The structural diagram of dSKN7-KanMX is shown below. Figure 1 As shown, the upstream and downstream homologous arms of SKN7 are homologous to the upstream and downstream sequences of the open reading frame (ORF) of the SKN7 gene on the chromosome, respectively. The KanMX expression frame refers to a complete DNA sequence with unidirectional loxP sequences at both ends, capable of expressing proteins that give the strain the G418 resistance characteristic. It includes the promoter TEFpromorter, the open reading frame KanR, and the terminator TEF terminator.

[0049] Example 2: Obtaining recombinant strains with the SKN7 gene knocked out

[0050] Transformants were obtained by transforming the starting strain BSGX001 with the dSKN7-KanMX fragment obtained in Example 1 and screened on YPD plates containing 200 μg / mL G418. The screened transformants were verified by extracting chromosomes and performing PCR (primers, PCR system, and conditions are detailed in Table 2). The transformant with a PCR product length of 1158 bp was the correct SNK7 knockout transformant BSGX001(skn7Δ), while transformants that did not knock out SNK7 could not produce PCR products. The DNA sequence at the SKN7 gene locus on the chromosome of transformant BSGX001(skn7Δ) was replaced by an expression frame sequence, thus preventing the transformant from expressing the Skn7 transcription factor.

[0051] Table 2

[0052]

[0053] Example 3: Test of the co-fermentation effect of glucose and xylose on mutant strains

[0054] The starting strain BSGX001 and the recombinant strain BSGX001(skn7Δ) were fermented in glucose-xylose co-fermentation medium, respectively. The cell growth, xylose utilization rate and ethanol accumulation of the starting strain and the recombinant strain during the fermentation process were compared.

[0055] The procedure is as follows: The starting strain and the recombinant strain were inoculated into SC-URA medium containing 20 g / L glucose and cultured at 30°C for 14 ± 2 h. The strains were then transferred to 300 mL Erlenmeyer flasks containing 50 mL of SC-URA medium. The carbon source was 20 g / L glucose. The initial OD... 600 The culture medium was 1.0, 200 rpm, and 30℃ for 12 h. After culture, the culture was centrifuged at 5500 rpm for 3 minutes, washed 1-2 times with sterile water, and resuspended in 50 mL of glucose-xylose cosaccharide fermentation medium. The initial OD was... 600 =2.5, fermented at 30℃ and 200rpm for 48h.

[0056] The cell count (expressed as OD) of the starting strain BSGX001 and the recombinant strain BSGX001(skn7Δ) during the fermentation process 600 The concentrations of glucose, xylose, and ethanol in the fermentation system were monitored (using the value as an evaluation index). Specifically, the HPLC analysis of glucose, xylose, and ethanol concentrations was performed using a Bio-Rad LC-20A system, and the chromatography column used was... HPX-87H (Bio-Rad) was analyzed using a Bio-Rad RID-10A differential detector. The mobile phase was 5 mmol / L H₂SO₄, the flow rate was 0.6 mL / min, and the column oven temperature was maintained at 45 °C. The xylose utilization rate was calculated using the following formula.

[0057]

[0058] Where r is the xylose specific utilization rate from sampling point m to sampling point n; A, B, and t are the xylose concentration, biomass concentration (i.e., cell dry weight), and t, respectively, at sampling points n, i, and m. 600 The fermentation broth was centrifuged, the cells were collected, dried, weighed, and the time was measured.

[0059] For detailed results, please see [link to results]. Figure 2 Where —□— represents the cell count of strain BSGX001 at various time points during fermentation, expressed as OD. 600 —■— represents the cell count of strain BSGX001(skn7Δ) at each time point during fermentation, expressed as OD600; —△— represents the glucose concentration of strain BSGX001 at each time point during fermentation; —▲— represents the glucose concentration of strain BSGX001(skn7Δ) at each time point during fermentation. —▼— Represents the xylose concentration of strain BSGX001 at various time points during fermentation; —○— Represents the xylose concentration of strain BSGX001(skn7Δ) at various time points during fermentation; —●— Represents the ethanol concentration of strain BSGX001(skn7Δ) at various time points during fermentation.

[0060] from Figure 2 As can be seen, the recombinant strain BSGX001(skn7Δ) exhibited better growth and xylose utilization than the original strain BSGX001 in glucose-xylose co-fermentation.

[0061] After 48 hours of fermentation, the cell mass of BSGX001(skn7Δ) was 18.5% higher than that of BSGX001; the xylose volume utilization rate (xylose utilization amount / time) throughout the entire stage was 15.2% higher than that of BSGX001, increasing from 0.22 g / L / h to 0.26 g / L / h; in the xylose utilization stage after glucose depletion, the xylose specific utilization rate of BSGX001(skn7Δ) was 8% higher than that of BSGX001, increasing from 0.113±0.002 g / L / h / g dry weight cells to 0.122±0.002 g / L / h / g dry weight cells; at the same time, the glucose utilization rate of BSGX001(skn7Δ) was comparable to that of the starting strain; in addition, compared with the starting strain, the ethanol accumulation of BSGX001(skn7Δ) after 48 hours was increased by 4%, from 12.4±0.1 g / L to 12.9±0.5 g / L. The above results indicate that knocking out the SKN7 gene, thereby eliminating the activity of the transcription factor Skn7, can significantly improve the xylose utilization capacity of Saccharomyces cerevisiae BSGX001.

[0062] Example 4

[0063] Using the methods described in Examples 1-2, the corresponding SKN7 gene knockout strain was prepared using Saccharomyces cerevisiae SDIC Sc E01 as the starting strain.

[0064] Using the method described in Example 3, Saccharomyces cerevisiae SDIC Sc E01 and its corresponding SKN7 gene knockout strain SDIC Sc E01 (skn7Δ) were fermented in glucose xylose co-fermentation medium for 48 hours. The cell growth, xylose utilization rate, and ethanol accumulation of the starting strain and the knockout strain during the fermentation process were compared.

[0065] Similar to BSGX001 and BSGX001(skn7Δ), the glucose utilization rate of SDIC Sc E01 is comparable to that of SDIC Sc E01(skn7Δ). The improvements of other parameters of SDIC Sc E01(skn7Δ) compared to SDIC Sc E01 are detailed in Table 3 below.

[0066] Table 3

[0067] Bacterial volume 16 Xylose volume utilization rate throughout the stage 15.7 Xylose utilization stage xylose specific utilization rate 7.8 Ethanol accumulation over 48 hours 5.2

[0068] The preferred embodiments of the present invention have been described in detail above; however, the present invention is not limited thereto. Within the scope of the inventive concept, various simple modifications can be made to the technical solutions of the present invention, including combinations of various technical features in any other suitable manner. These simple modifications and combinations should also be considered as the content disclosed in the present invention and are all within the protection scope of the present invention.

Claims

1. A brewing yeast with improved xylose utilization efficiency, characterized in that, The brewing yeast is selected from knockout yeast. SKN7 Saccharomyces cerevisiae CGMCC No. 17264 and / or knockout genes SKN7 The gene of Saccharomyces cerevisiae GDMCC No:62491.

2. A method for improving the xylose utilization growth ability of Saccharomyces cerevisiae and / or improving the xylose utilization efficiency of Saccharomyces cerevisiae in a glucose-xylose co-fermentation system, characterized in that, The method includes knocking out brewer's yeast. SKN7 The gene, wherein the brewer's yeast is selected from brewer's yeast CGMCC No.17264 and / or GDMCC No:62491.

3. The application of the brewing yeast of claim 1, and / or the method of claim 2, in ethanol production.

4. The brewing yeast of claim 1, and / or the method of claim 2, in the production of ethanol by fermentation with a substrate containing lignocellulose hydrolysate.

5. A method for producing ethanol, characterized in that, The method includes fermenting a substrate containing glucose and xylose using brewer's yeast and then knocking out the brewer's yeast. SKN7 The gene, wherein the brewer's yeast is selected from brewer's yeast CGMCC No. 17264 and / or GDMCC No: 62491.

6. The method according to claim 5, wherein, The substrate includes a hydrolysate of lignocellulose raw material.

7. The method according to claim 5, wherein, The substrate also contains at least one of mannose and galactose as a carbon source.

8. The method according to claim 5, wherein, The weight ratio of glucose to xylose in the substrate is 1:0.5-5.

9. The method according to claim 5, wherein, The fermentation conditions include initial OD 600 =0.5-5, speed 100-300rpm, temperature 25-35℃.

Images