CN113462587B

CN113462587B - Saccharomycopsis strain for high yield of amylase and preparation method and application thereof

Info

Publication number: CN113462587B
Application number: CN202110849810.4A
Authority: CN
Inventors: 刘锐; 刘功良; 廖志诚; 卢方正; 赵翾
Original assignee: Zhongkai University of Agriculture and Engineering
Current assignee: Zhongkai University of Agriculture and Engineering
Priority date: 2021-07-27
Filing date: 2021-07-27
Publication date: 2022-10-25
Anticipated expiration: 2041-07-27
Also published as: CN113462587A

Abstract

The invention relates to the technical field of microorganisms, and particularly relates to a sacculus-covering yeast strain for high yield of amylase and a preparation method and application thereof. The invention improves the strain by an ultraviolet mutagenesis method, selects excellent strains after each mutagenesis for repeated mutagenesis, and obtains a mutant strain sacculus-covering yeast strain lglY1M with stable hereditary characters after multiple separation and purification. Compared with the enzyme activity of fermentation liquor before mutagenesis and after mutagenesis through experiments, the result proves that the highest enzyme activity of the mutagenized strain reaches 48.0U/L, the highest enzyme activity of the original strain before mutagenesis is 15.9U/L, the enzyme activity is improved by two times, and the enzyme activity is still better when the alcohol degree is up to 15%. The sacculus-covering yeast strain provided by the invention has the advantages that the amylase yield and the amylase activity are obviously improved, the performance is stable, and the development prospect in the industrial application of the amylase is wide.

Description

Saccharomycopsis strain for high yield of amylase and preparation method and application thereof

Technical Field

The invention relates to the technical field of microorganisms, in particular to a high-amylase-yield sacculus-covering yeast strain and a preparation method and application thereof.

Background

Amylase is a glycoside hydrolase, the substrate of which is starch, and can decompose starch into small molecular substances such as dextrin, maltose, glucose and the like, and the amylase can be divided into alpha-amylase, beta-amylase, saccharifying enzyme, isoamylase and the like according to different action modes. The amylase is the most extensive hydrolase in industrial production and application, can account for about one fourth of market share, and has great use value in the directions of fermentation, food, medicine, spinning, cleaning and the like. Microbial amylases have been very successful alternatives to chemically hydrolyzing starch since 1894 the first commercial amylase was derived from fungi. Due to the wide application range and great yield demand of amylase, the amylase is of great importance in the fermented food industry.

The sacculus-covering yeast is also called endospora, has the characteristic of rapidly producing amylase, acid protease and beta-glucosidase in large quantity, can produce alcohol by utilizing carbohydrates such as sucrose, cellobiose, soluble starch and the like, and widely exists in various distillers yeast, such as Fendaqu, red yeast rice, luzhou-flavor liquor Daqu, xiaoqu liquor Daqu drug, maotai-flavor Daqu and fermented grains in the early fermentation stage, so the sacculus-covering yeast is considered to have larger application potential in the brewing industry.

The current research on the Saccharopolyspora sinensis focuses on the extracellular hydrolases secreted by it, including beta-glucosidase, amylase, protease, etc. However, the amylase activity produced by the saccharomycetes with the envelope covering and screened from the nature is low, the requirement of industrial production cannot be met, and the wide application of the amylase is severely restricted.

Disclosure of Invention

The invention aims to provide a high-amylase-yield saccule-coated yeast strain and a preparation method thereof aiming at the defects of the prior art, wherein the highest enzyme activity of the strain is improved by more than two times compared with that of an original strain before mutagenesis, and the strain has the advantages of high enzyme production activity and good stability.

The invention also aims to provide the application of the high-yield amylase-producing envelope-covering yeast strain in amylase production.

The purpose of the invention is realized by the following technical scheme:

provides a sacculus-coated yeast strain for high yield of amylase, which is sacculus-coated yeast lglY1M, is classified and named as Saccharomyces fibuligera lgy 1M, is deposited in Guangdong province microorganism strain collection center, and has the preservation number of GDMCC NO:61791, preservation date 2021, 7 months and 8 days.

The invention also provides a preparation method of the high-yield amylase-producing saccule-coated yeast strain, which comprises the following steps:

step a, ultraviolet mutagenesis:

a1 Carrying out plate-line drawing to culture strains, culturing for 2d in a constant-temperature incubator at 37 ℃, then inoculating the strains into a 5ml test tube enrichment medium in an aseptic operation, and culturing for 12h;

a2 Pouring the enriched and cultured bacterial suspension into a sterilized flat plate, placing the flat plate on a magnetic stirrer for ultraviolet mutagenesis, adding 5ml of enriched culture medium into the mutagenized bacterial suspension after the ultraviolet mutagenesis is finished, then completely wrapping the bacterial suspension by using tinfoil paper, and placing the bacterial suspension into a constant-temperature incubator at 37 ℃ for dark reaction for 1h;

a3 Shaking the bacterial suspension after the dark reaction, sucking a certain amount of bacterial suspension for gradient dilution, then uniformly coating the bacterial suspension on a flat plate, and placing the flat plate in a constant-temperature incubator at 37 ℃ for culturing for 48 hours;

a4 Carrying out iodine fumigation on the colonies cultured for 48 hours by using an iodine fumigation method, selecting the colonies with a large ratio of the area of the transparent ring to the area of the colonies by using a double-ring analysis method for enrichment culture, and then carrying out mutagenesis again;

a5 Repeatedly mutagenizing for multiple times until the ratio of the area of a transparent circle of a bacterial colony to the area of the bacterial colony before and after mutagenesis is obviously different, and stopping mutagenesis;

step b, separation and purification:

b1 Carrying out enrichment culture on the excellent strains screened out after mutagenesis for a certain time, then carrying out gradient dilution and coating, and placing in a constant-temperature incubator at 37 ℃ for culture for 48h;

b2 Carrying out iodine fumigation on the colonies cultured for 48 hours by using an iodine fumigation method, selecting excellent strains by using a double-circle analysis method, carrying out enrichment culture, plate coating and iodine fumigation in sequence until the finally coated strains have stable properties, namely the ratio of the transparent circle area of the colonies before and after mutagenesis to the colony area is obviously different, and stopping separation and purification to obtain the microcapsule coated saccharomycete with high amylase yield.

In the technical scheme, the enrichment medium is prepared from the following components:

10g of soluble starch, 10g of yeast extract powder, 20g of peptone and 1000ml of water.

In the above technical scheme, in the step a2, the ultraviolet mutagenesis conditions are as follows: the ultraviolet irradiation time is 20-60 min, and the ultraviolet irradiation distance is 30cm.

In the above technical scheme, in step a5, the number of times of repeated mutagenesis is 6 to 12.

The invention also provides application of the sacculus-covering yeast strain for producing the amylase at high yield in the production of the amylase.

The invention has the beneficial effects that:

the invention improves the strain by an ultraviolet mutagenesis method, selects excellent strains after each mutagenesis for repeated mutagenesis, selects the strains for separation and purification after the area of a transparent ring is obviously increased after the mutagenesis, and obtains a mutant strain cyst-coating yeast strain lglY1M with stable hereditary characters after multiple times of separation and purification. Compared with the enzyme activity of fermentation liquor before mutagenesis and after mutagenesis through experiments, the result proves that the highest enzyme activity of the mutagenized strain reaches 48.0U/L, the highest enzyme activity of the original strain before mutagenesis is 15.9U/L, the enzyme activity is improved by two times, amylase produced by the strain still has better enzyme activity when the alcohol degree is up to 15%, and the enzyme activity is kept above 80%. Therefore, the sacculus-covering yeast strain provided by the invention has the advantages that the amylase yield and the amylase activity are obviously improved, the performance is stable, the requirement of industrial production can be better met, and the development prospect in industrial application of amylase is wide.

Drawings

The invention is further illustrated by means of the attached drawings, but the embodiments in the drawings do not constitute any limitation to the invention, and for a person skilled in the art, other drawings can be obtained on the basis of the following drawings without inventive effort.

FIG. 1 is a schematic diagram showing the sizes of transparent circles on the same plate of the mutagenized strain and the non-mutagenized strain of this example.

FIG. 2 is a graph showing a glucose standard curve in the present example.

FIG. 3 is a graph showing comparison of enzyme activities of the mutagenized strain and the non-mutagenized strain of this example.

FIG. 4 is a graph showing the growth curves and enzyme activities of the mutagenized strain and the non-mutagenized strain of this example in comparison.

FIG. 5 is a graph showing the effect of temperature on the amylase activity of Saccharomyces cerevisiae.

FIG. 6 is a diagram showing the effect of alcohol content on amylase activity of Saccharomyces cerevisiae.

FIG. 7 is a graph showing the effect of pH on amylase activity produced by Saccharomycopsis fibuligera.

Detailed Description

The invention is further described in connection with the following examples.

The high-amylase-yield Saccharopolyspora sinensis strain lglY1M of the embodiment is classified and named as Saccharomyces fibuligera lgy 1M, and is deposited in Guangdong province culture collection center, with the deposition address: china Guangdong, with the preservation number GDMCC NO:61791, preservation date 2021, 7 months and 8 days.

1. The experimental method comprises the following steps:

1. determination of ultraviolet mutagenesis conditions:

a. carrying out plate-drawing method to culture strains, culturing for 2d in a constant-temperature incubator at 37 ℃, then inoculating the strains into 5ml of test tube enrichment medium by aseptic operation, and culturing for 12h;

b. sucking 100 mul from the bacteria suspension liquid for gradient dilution, sucking 100 mul to a flat plate for even coating by a coater, and placing the flat plate in a constant-temperature incubator at 37 ℃ for culture;

c. pouring the bacterial suspension subjected to enrichment culture into a sterilized flat plate, placing the flat plate on a magnetic stirrer, performing ultraviolet mutagenesis in an ultra-clean workbench, opening a cover of the flat plate during mutagenesis, and placing a stirrer into the bacterial liquid to enable the bacterial liquid to uniformly irradiate ultraviolet rays;

d. setting different groups with mutagenesis time of 20min, 30min,40 min, 50min and 60min respectively for mutagenesis, adding 5ml of enrichment culture medium into the mutagenized bacterial suspension after ultraviolet mutagenesis is finished, then completely wrapping the bacterial suspension by using tinfoil paper, and putting the bacterial suspension into a constant-temperature incubator at 37 ℃ for dark reaction for 1h;

e. shaking the bacterial suspension after the dark reaction, sucking 100 mul for gradient dilution, sucking 100 mul to a flat plate, coating evenly by using a coater, and placing in a constant-temperature incubator at 37 ℃ for culturing for 48h;

f. the number of colonies before and after mutagenesis was counted, and then the lethal amount was calculated, and the results are shown in FIG. 1.

As can be seen from figure 1, the lethal dose of the saccule-coated saccharomycete reaches over 99 percent when the ultraviolet radiation is used for mutagenesis for 20-60 min and the distance is 30cm. Therefore, the envelope saccharomycete is induced by ultraviolet irradiation within the range of the irradiation distance and the irradiation time, so that the strain is optimized and improved.

2. Ultraviolet mutagenesis:

a2 The bacterial suspension is poured into a sterilized flat plate, and placed on a magnetic stirrer for ultraviolet mutagenesis, wherein the ultraviolet irradiation time is 20-60 min, and the ultraviolet irradiation distance is 30cm. After ultraviolet mutagenesis is finished, adding 5ml of enrichment medium into the mutagenized bacterial suspension, then completely wrapping the bacterial suspension by using tinfoil paper, and putting the bacterial suspension into a constant-temperature incubator at 37 ℃ for dark reaction for 1 hour;

a4 Performing iodine fumigation on the colonies cultured for 48 hours by using an iodine fumigation method, selecting the colonies with a large ratio of the area of the transparent ring to the area of the colonies by using a double-ring analysis method to perform enrichment culture, and then performing mutagenesis again;

a5 Repeatedly mutagenizing for multiple times until the ratio of the transparent circle area of the colony to the colony area before and after mutagenesis is obviously different, and stopping mutagenesis.

In this example, mutagenesis was performed for one month, and 3 to 4 strains among 300 to 500 colonies were selected for each mutagenesis as the most excellent strains among the batch mutagenesis, which were then subjected to enrichment culture, and then to repeated mutagenesis. The ratio of clearing circle area to colony area had increased to 8.3, 9.2, 7.5 and 7.3 after 6 mutagenizations. Therefore, it was confirmed that the mutagenesis of the strain could be stopped, and then several colonies which were most excellent were selected and isolated and purified.

3. Separation and purification:

b2 Carrying out iodine fumigation on the colonies cultured for 48 hours by using an iodine fumigation method, selecting excellent strains by using a double-circle analysis method, and then sequentially carrying out enrichment culture, plate coating and iodine fumigation until the characteristics of the finally coated strains are stable, namely the ratio of the transparent circle area of each colony to the colony area is stable and has obvious difference from the ratio of the transparent circle area of the colony to the colony area before mutagenesis, and then stopping separation and purification.

In the above steps, the enrichment medium is prepared from 10g of soluble starch, 10g of yeast extract powder, 20g of peptone and 1000ml of water, and is sterilized at 120 ℃ for 15min.

2. Verification and analysis of mutant strain enzyme-producing activity

1. Plate verification:

the mutagenized and non-mutagenized strains were plated on the same plate for 48h and then iodine-fumigated for comparison of the size of the clearing circle, the results are shown in FIG. 1.

As can be seen from FIG. 1, the ratio of the transparent circle area of the non-mutagenized strain to the colony area is 4.9, 5.3, 5.4, 4.7, 5.1 and 4.8, and the ratio of the transparent circle area of the mutagenized strain to the colony area is 6.8, 5.0, 6.0, 5.7, 6.6, 6.0, 5.8 and 5.9. This shows that the characters of the mutant strain are basically stable after multiple times of ultraviolet mutagenesis and separation and purification.

2. Drawing a glucose standard curve:

accurately weighing 0.1g of anhydrous glucose, dissolving with deionized water, diluting to 100ml of constant volume for later use, accurately transferring liquid to a 25ml colorimetric tube according to data in table 1, and taking a No. 7 tube as a blank control group. Transferring the solution, placing the colorimetric tube in boiling water bath for 7min, then placing in ice water bath, rapidly cooling for 2min, metering volume to 25ml, performing color reaction for 30min, and measuring light absorption value at 500nm wavelength. The absorbance was measured at a wavelength of 500nm using DNS-aqueous solution, tube No. 7, as a blank. The glucose standard curve was plotted with the glucose mass concentration (x) as the abscissa and the absorbance value (y) as the ordinate, and the result is shown in fig. 2.

TABLE 1 glucose solution concentration

As shown in fig. 2, the linear regression equation for the standard curve is: y =0.9903x-0.0332,R ² The linear relation of the mass concentration of the glucose is 0.1-1.0, and the method for measuring the activity of the amylase by measuring the concentration of the reducing sugar has better accuracy.

3. And (3) measuring the enzyme activity of the fermentation liquor:

a. weighing 80g of ammonium sulfate, heating to dissolve with 100ml of water, heating to about 60 ℃, and then placing the solution in a refrigerator at 4 ℃ to prepare a saturated ammonium sulfate solution for later use.

b. Inoculating the separated and purified strains into a 5ml test tube enrichment medium for enrichment culture, and simultaneously inoculating strains which are not subjected to mutagenesis into the 5ml test tube enrichment medium for enrichment culture for later use.

c. The two strains which were subjected to enrichment culture for 24 hours were added to 500ml Erlenmeyer flasks containing 100ml of fermentation medium, and then placed on a shaker for culture (37 ℃,120 r/min).

d. Removing the bacterial suspension from the fermentation broth at regular intervals, and measuring the absorbance of the bacterial suspension at a wavelength of 560 nm. Meanwhile, 1.6ml of the bacterial suspension is transferred into a 4ml centrifuge tube, then 2.4ml of saturated ammonium sulfate solution is added, and the mixture is placed in a refrigerator at 4 ℃ for salting out for 4 hours. Several preparations were required for both mutagenized and unmutagenized bacteria.

e. After salting out for 4h, centrifuging (10 min, 12000r/min), pouring out the supernatant, redissolving with 0.8ml deionized water for each tube, and repeatedly washing the tube wall of the centrifuge tube during redissolving.

f. And (3) mixing the redissolved solution according to the proportion of 1:2 and 1mg/ml starch solution, adding into a test tube, inactivating enzyme, reacting in 37 deg.C water bath for 30min, rapidly inactivating enzyme in boiling water bath for 10min, cooling in ice water bath for 2min, and centrifuging (10min, 12000r/min).

g. After centrifugation, 1ml of supernatant is transferred into a 25ml colorimetric tube, then 1.5ml of DNS developer is transferred and placed into boiling water bath for 7min, ice water bath for 2min, constant volume is carried out to 25ml, shaking is carried out, color reaction is carried out for 30min, the light absorption value is measured under the condition of 500nm wavelength, and DNS-aqueous solution is used as a blank control group.

h. And calculating the enzyme activity.

The comparison of the enzyme activities and growth curves of the mutagenized bacteria and the unmutagenized bacteria are shown in FIGS. 3 and 4, respectively. The results indicate that the UV-mutagenized strains grew at a slower rate than the non-mutagenized strains. The time for producing amylase by the saccharomycete covered with capsules is started after 12 hours of culture of fermentation liquor, and the growth of strains is mainly carried out in the early stage, so that no amylase is produced or the quantity of the produced amylase is very small. After 24 hours of fermentation broth culture, the enzyme activity can be measured, and the activity of the amylase produced by the mutagenic bacteria after 24 hours of culture is almost the same as that of the amylase produced by the non-mutagenic bacteria; after 48 hours of fermentation broth culture, the activity of amylase produced by the mutagenic bacteria is more than 5 times that of the non-mutagenic bacteria, and the difference of the enzyme activity is the largest; after the fermentation broth is cultured for 72 hours, the activity of the amylase produced by the mutagenic bacteria and the amylase produced by the non-mutagenic bacteria reach the highest point, and at the moment, the activity of the amylase produced by the mutagenic bacteria is more than 3 times that of the amylase produced by the non-mutagenic bacteria; after 96 hours of fermentation broth culture, the activity of amylase produced by the mutagenic bacteria and the non-mutagenic bacteria begins to decrease, especially the activity of the amylase of the mutagenic bacteria is sharply decreased, and the activity of the amylase produced by the mutagenic bacteria and the non-mutagenic bacteria is almost the same. Therefore, experiments show that the growth speed of the mutagenic bacteria is slower than that of the non-mutagenic bacteria, but the produced amylase activity is that the highest enzyme activity of the mutagenic strain reaches 48.0U/L, while the highest enzyme activity of the original strain before mutagenesis is 15.9U/L, and the enzyme activity is improved by two times.

4. Influence of temperature on enzyme activity:

a. and inoculating the separated and purified strains into a 5ml test tube enrichment medium for enrichment culture, and simultaneously inoculating strains which are not subjected to mutagenesis into the 5ml test tube enrichment medium for enrichment culture for later use.

b. The two strains which are subjected to enrichment culture for 24 hours are respectively added into a 500ml triangular flask of 100ml fermentation medium, and then placed on a shaking table for culture for 72 hours. (37 ℃,120 r/min).

c. Transferring 10ml of the bacterial suspension and 15ml of ammonium sulfate saturated solution into a 50ml centrifuge tube, placing in a refrigerator at 4 ℃ for salting out for 4h, then centrifuging (30min, 4000 r/min), pouring out the supernatant, transferring 5ml of ionized water for redissolution,

d. and (3) mixing the redissolved solution according to the proportion of 1:2 and 1mg/ml starch solution, adding into a test tube, inactivating enzyme, reacting in water bath at gradient temperature for 30min, rapidly inactivating enzyme in water bath for 10min, cooling in ice water bath for 2min, and centrifuging (10min, 12000r/min).

e. After centrifugation, 1ml of supernatant is transferred into a 25ml colorimetric tube, then 1.5ml of DNS developer is transferred and placed into boiling water bath for 7min, ice water bath for 2min, constant volume is carried out to 25ml, shaking is carried out, color reaction is carried out for 30min, the light absorption value is measured under the condition of 500nm wavelength, and DNS-aqueous solution is used as a blank control group. The results are shown in FIG. 5.

As can be seen from FIG. 5, the amylase produced by the saccharomycete with envelope covering and membrane covering has the highest reaction enzyme activity at 42 ℃ within 30min, has the enzyme activity of more than 80% at the temperature of 32-52 ℃, has little influence on the enzyme activity at the temperature of less than 42 ℃, and has the relative enzyme activity of more than 80% at room temperature; when the temperature is up to 57 ℃, the enzyme activity is sharply reduced, mainly because the temperature is too high, partial amylase is denatured and loses activity. Therefore, the amylase produced by the sacculus complex membrane yeast is suitable for catalytic hydrolysis reaction at a lower temperature and is not suitable for catalytic hydrolysis reaction at a high temperature of more than 57 ℃.

5. Influence of alcoholic strength on enzyme activity:

b. The two strains which are subjected to enrichment culture for 24 hours are respectively added into a 500ml triangular flask of 100ml fermentation medium, and then are placed on a shaking table for culture for 72 hours. (37 ℃,120 r/min).

c. Transferring 10ml of the bacterial suspension and 15ml of ammonium sulfate saturated solution into a 50ml centrifuge tube, placing the centrifuge tube in a refrigerator at 4 ℃ for salting out for 4h, then centrifuging (30min, 4000 r/min), pouring out supernatant, and transferring 5ml of solution with different alcoholic strength for redissolution.

d. And (3) mixing the redissolved solution according to the proportion of 1:2 and 1mg/ml starch solution, adding into a test tube, inactivating enzyme, reacting in water bath at 37 deg.C for 30min, inactivating enzyme in boiling water bath for 10min, cooling in ice water bath for 2min, and centrifuging (10min, 12000r/min).

e. After centrifugation, 1ml of supernatant is transferred into a 25ml colorimetric tube, then 1.5ml of DNS developer is transferred and placed into boiling water bath for 7min, ice water bath for 2min, constant volume is carried out to 25ml, shaking is carried out, color reaction is carried out for 30min, the light absorption value is measured under the condition of 500nm wavelength, and DNS-aqueous solution is used as a blank control group. The results are shown in FIG. 6.

As can be seen from FIG. 6, the amylase produced by the saccharomycete with coated capsules has the highest enzyme activity within 30min of reaction time when the alcohol content is 5%; when the alcoholic strength is lower than 15%, the influence on the enzyme activity of the amylase is small, and the enzyme activity can reach more than 80%; when the alcoholic strength reaches 20%, the enzyme activity is seriously reduced, and the enzyme activity is reduced because partial protein is denatured due to overhigh alcohol concentration. Therefore, the amylase produced by the saccharomycete with the sacculus complex membrane has higher catalytic hydrolysis reaction efficiency when the alcoholic strength is lower than 15%, and the catalytic reaction efficiency is reduced when the alcoholic strength is higher than 20%.

6. Influence of pH on enzyme Activity:

c. Transferring 10ml of the bacterial suspension and 15ml of ammonium sulfate saturated solution into a 50ml centrifuge tube, placing the centrifuge tube in a refrigerator at 4 ℃ for salting out for 4h, then centrifuging (30min, 4000 r/min), pouring out supernatant, and transferring 5ml of solution with different pH values for redissolution.

d. And (3) mixing the redissolved solution according to the proportion of 1:2 and 1mg/ml starch solution, adding into a test tube, inactivating enzyme to serve as a control group, reacting in a water bath at 37 ℃ for 30min, rapidly inactivating enzyme in a boiling water bath for 10min, cooling in an ice water bath for 2min, and centrifuging (10min, 12000r/min).

e. Centrifuging, transferring 1ml of supernatant into a 25ml colorimetric tube, transferring 1.5ml of DNS developer, putting the DNS developer into boiling water for boiling water bath for 7min, performing ice water bath for 2min, fixing the volume to 25ml, shaking up, performing color reaction for 30min, measuring the light absorption value under the condition of 500nm wavelength, and taking DNS-aqueous solution as a blank control group. The results are shown in FIG. 7.

As can be seen from FIG. 7, the amylase produced by the saccharomycete with coated capsules reacts for 30min under different pH values, the change of the enzyme activity is relatively large, and the enzyme activity is highest under the condition of neutral pH value; under the acidic condition of pH 4-6, the enzyme activity is more than 80 percent, and the influence on the enzyme activity is little; when the pH is in a weak alkali environment, the enzyme activity is sharply reduced. Therefore, the amylase produced by the sacculus complex membrane yeast is suitable for catalytic hydrolysis reaction under acidic and neutral conditions with the pH value of 4-7 and is not suitable for catalytic hydrolysis reaction under alkaline conditions.

Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention, and not for limiting the protection scope of the present invention, although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims

1. A high-amylase-yield sacculus-covered yeast strain is characterized in that: the strain is a sacculus complex membrane yeast lglY1M, is classified and named as Saccharomyces fibuligera lgy 1M, is preserved in Guangdong province microbial strain preservation center, and has the preservation number of GDMCC NO:61791, with a preservation date of 2021, 7 months and 8 days.

2. The use of an amylase-producing strain of Saccharopolypermannide yeast according to claim 1 for the production of amylase.