Temperature-resistant Jutanberg zebnella yeast and application of temperature-resistant Jutanberg zebnella yeast

Abstract

The invention relates to the technical field of microorganisms, and discloses a temperature-resistant Jutanberg's heldnella yeast and an application thereof, and the Jutanberg's heldnella yeast is Jutanberg's heldnella yeast 413, and the temperature-resistant Jutanberg's heldnella yeast is Jutanberg's heldnella yeast 413. The invention relates to a Jerbudzea jadinii strain which is characterized in that the Jerbudzea jadinii strain 413 is preserved in Guangdong Province Microbial Culture Collection Center on November 11, 2024, the preservation address is the 5th floor of the building 59, No. 100 Courtyard, Xianlie Middle Road, Guangzhou City, the preservation number is GDMCC NO: 65456, and the microorganism classification is named as the Jerbudzea jadinii 413. According to the present invention, the lindnella gedinieri has good temperature resistance, and can be fermented at the fermentation temperature of 28-38 DEG C so as to obtain the protein feed with the high protein content;

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to a temperature-resistant yeast Jardine Seiberrinna and an application thereof. Background Art

[0002] my country is very short of protein resources, especially feed protein. my country's annual feed protein consumption in the livestock industry is about 47.54 million tons, of which 69% is imported. The shortage of protein feed has seriously affected the development of my country's livestock and animal husbandry industries, so more and more extensive protein feed resources are needed to invest in livestock production.

[0003] Lindnerella jardiniseiber Cyberlindenera jadinii ) is one of the excellent strains for high-yield bacterial protein in microbial fermentation feed. It is rich in vitamin B and protein, and has the effects of regulating the balance of animal intestinal microecology, improving feed digestibility, and enhancing animal immunity. It has the advantages of short culture cycle, good safety and good fermentation performance. It can use industrial by-products such as molasses, starch factory waste liquid and distiller's grains as carbon sources, and urea or nitrate as nitrogen sources to grow and ferment feed. However, during the fermentation and protein conversion process of existing strains, the fermentation environment temperature will rise, often higher than the set fermentation temperature, which inhibits the activity and metabolic process of Jedinseiberlindner's yeast in the fermentation environment. Therefore, it is necessary to select strains with better temperature resistance. Summary of the invention

[0004] In order to solve the above technical problems, the present invention provides a temperature-resistant Jardine Seiberlindera yeast and its application. The Jardine Seiberlindera yeast in the present invention has good temperature resistance and can be fermented at a fermentation temperature of 28°C-38°C to obtain a protein feed with a higher protein content.

[0005] Jardineseberlindner's yeast 413 was deposited in the Guangdong Microbial Culture Collection Center (GDMCC) on November 11, 2024. The deposit address is 5th Floor, Building 59, No. 100, Xianlie Middle Road, Guangzhou. The deposit number is GDMCC NO: 65456. The microbial classification is named Jardineseberlindner's yeast Cyberlindenera jadinii 413.

[0006] The present invention provides a temperature-resistant Jardineseberlindner yeast, wherein the Jardineseberlindner yeast is Jardineseberlindner yeast 413; The Jadinseberlindner yeast 413 has been deposited in the Guangdong Microbial Culture Collection Center (GDMCC) on November 11, 2024, with the deposit address being 5th Floor, Building 59, No. 100, Xianlie Middle Road, Guangzhou, with the deposit number being GDMCC NO: 65456, and the microbial classification is named Jadinseberlindner yeast Cyberlindenera jadinii 413.

[0007] Furthermore, the culture temperature of the Saccharomyces jedinseberlindera 413 is 28°C-38°C.

[0008] Furthermore, the culture temperature of the Saccharomyces jedinseberlindera 413 is 35°C-38°C.

[0009] The present invention also provides the use of the Jardine Seiberlindner yeast in preparing protein feed by fermenting corn.

[0010] Further, the corn syrup was liquefied, enzymatically hydrolyzed to reduce viscosity, saccharified, and nitrogen source was added. The yeast solution of Saccharomyces cerevisiae was inoculated into the fermentation environment at 28°C-38°C and a ventilation volume of 1m 3 / h-1.5m 3 / h, ferment for 28h-32h, after the fermentation is completed, separate the solid and liquid, and dry the solid to obtain protein feed; The Lindnerella jardiniseiber is Lindnerella jardiniseiber 413.

[0011] Furthermore, the mass ratio of the bacterial liquid of the Saccharomyces jadinseberlindnerii to the corn steep liquor in the fermentation environment is 5%.

[0012] Furthermore, the specific method of the liquefaction treatment includes: adjusting the temperature and pH value of the corn slurry, and adding liquefaction enzyme treatment.

[0013] Further, the temperature of the corn steep liquor is adjusted to 91°C-97°C.

[0014] Furthermore, the pH value is 5-6.

[0015] Furthermore, the liquefaction enzyme includes a liquefaction enzyme with an activity of 10,000-20,000 U / g.

[0016] Furthermore, the mass concentration of the liquefaction enzyme in the corn steep liquor is 0.2%.

[0017] Those skilled in the art should understand that stirring may be performed during the liquefaction process to facilitate the liquefaction process.

[0018] Furthermore, an iodine test is performed on the corn syrup during the liquefaction process. When the substrate of the liquefaction process reacts with the iodine solution and changes from blue to orange during the iodine test, the liquefaction process is completed.

[0019] Furthermore, the specific method of reducing viscosity by enzymatic hydrolysis includes: adding enzymatic hydrolyzate to the liquefied corn steep liquor, adjusting pH, and heating in a water bath.

[0020] Furthermore, the enzymatic hydrolysate includes one or more of cellulase and pectinase.

[0021] Furthermore, the cellulase has an activity of 400,000-500,000 U / g.

[0022] Furthermore, the pectinase has an activity of 10,000-20,000 U / g.

[0023] Furthermore, the amount of the enzymatic hydrolysate added to the corn steep liquor after liquefaction is 0.2% by weight.

[0024] Furthermore, the pH is adjusted to 4-5.

[0025] Furthermore, the water bath heating temperature is 50°C-55°C.

[0026] It should be understood by those skilled in the art that stirring may be performed during the process of enzymatic viscosity reduction, thereby facilitating the viscosity reduction process.

[0027] Furthermore, the viscosity of the corn steep liquor during the enzymatic viscosity reduction process is measured, and when the viscosity is lower than 2000 mPa.s, the enzymatic viscosity reduction is completed.

[0028] Furthermore, the specific method of the saccharification treatment includes: adding saccharifying enzyme to the corn steep liquor after enzymatic hydrolysis and viscosity reduction, adjusting pH, and heating in a water bath.

[0029] Furthermore, the added mass of the saccharifying enzyme in the corn steep liquor after enzymatic hydrolysis and viscosity reduction is 0.6%.

[0030] Furthermore, the saccharifying enzyme includes a saccharifying enzyme with an activity of 40,000-50,000 U / g.

[0031] Furthermore, the pH is adjusted to 4-5.

[0032] Furthermore, the water bath heating temperature is 55°C-60°C.

[0033] Those skilled in the art should understand that stirring may be performed during the saccharification process to facilitate the saccharification process.

[0034] Furthermore, the presence of dextrin in the corn slurry during the saccharification process is detected by an anhydrous ethanol method, and the saccharification process is terminated when no dextrin is present in the corn slurry during the saccharification process.

[0035] Furthermore, the nitrogen source includes ammonium sulfate.

[0036] Furthermore, the nitrogen source is added in an amount of 1%-2% to the corn steep liquor after saccharification.

[0037] Furthermore, during fermentation, the gas introduced is air. Furthermore, the preparation of the corn slurry is specifically as follows: corn and water are pulped, and the mass ratio of corn to water is 1:1.25.

[0038] Furthermore, the method for preparing the bacterial liquid of Saccharomyces jadiniensis 413 includes: placing Saccharomyces jadiniensis 413 in a shake flask culture medium and culturing at 28° C.-38° C. for 24 h-28 h.

[0039] Furthermore, the formula of the shake flask culture medium is: yeast extract, glucose, magnesium sulfate, and dipotassium hydrogen phosphate; the mass concentration ratio of the yeast extract, glucose, magnesium sulfate, and dipotassium hydrogen phosphate in the shake flask culture medium is (8.5g / L-10.5g / L):(18g / L-21g / L):(1g / L-1.5g / L):(1.3g / L-1.5g / L).

[0040] Furthermore, those skilled in the art should understand that the purpose of solid-liquid separation is to obtain the fermentation product, and those skilled in the art can select different methods according to actual conditions as long as they can achieve the purpose. As an example, centrifugation is used at 8000 rpm for 6 minutes.

[0041] Furthermore, the drying temperature is 55°C-65°C.

[0042] Furthermore, the powder needs to be crushed after drying, and those skilled in the art can select the crushing mesh size according to actual conditions.

[0043] The embodiments of the present invention have the following technical effects: 1. The Jardine Seberlindner yeast of the present invention has good temperature resistance, can be better fermented in an environment of 28°C-38°C, and can successfully use corn to obtain a protein feed.

[0044] 2. The Jadinseberlindner yeast of the present invention is applied to fermented corn slurry, and corn is added with water to prepare a slurry. The addition of an appropriate amount of water can help the nutrients in the corn to dissolve and leach better, which is beneficial to liquefaction, enzymatic viscosity reduction and saccharification. First, liquefaction is carried out, and the material-liquid ratio is optimized in the liquefaction process. First, the fermentation substrate has a certain fluidity, which is beneficial to the efficiency of the enzyme action in the subsequent enzymatic viscosity reduction process; second, the starch in the fermentation substrate is gelatinized at high temperature and broken into short chains to produce dextrin and sugars, which are more easily saccharified by saccharifying enzymes during the saccharification process. Enzymatic viscosity reduction decomposes the crude fiber in the fermentation substrate into monosaccharides and oligosaccharides through the joint action of cellulase and pectinase, and can increase the surface area and porosity of the fermentation substrate, increase the contact area and utilization rate of the substrate, and reduce the viscosity of the substrate, so that the subsequent saccharifying enzyme action is more efficient and sufficient. The saccharification treatment further decomposes the dextrin and oligosaccharides in the fermentation system into monosaccharides that can be used by yeast, providing sufficient carbon sources for yeast fermentation and proliferation. Under this process, the fermentation time is greatly shortened, and a cycle can be completed in 30 hours. Third, the fermentation raw material corn pulp comes from whole ears of corn, which improves the utilization rate of corn resources and reduces the waste of corn cob resources.

[0045] 3. The method of the present invention is simple to operate, has a short production cycle and low production cost. The present invention converts corn into protein through fermentation, which not only fills the demand gap for protein feed in my country, but also is conducive to the recycling of agricultural waste. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0047] Figure 1 It is the test result of the viable bacteria count provided by the embodiments and comparative examples of the present invention.

[0048] Figure 2 These are the protein content test results provided by the embodiments and comparative examples of the present invention. DETAILED DESCRIPTION

[0049] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work belong to the scope of protection of the present invention.

[0050] In a first aspect, some embodiments of the present invention provide a thermotolerant yeast, wherein the yeast is yeast 413; The Jadinseberlindner yeast 413 has been deposited in the Guangdong Microbial Culture Collection Center (GDMCC) on November 11, 2024, with the deposit address being 5th Floor, Building 59, No. 100, Xianlie Middle Road, Guangzhou, with the deposit number being GDMCC NO: 65456, and the microbial classification is named Jadinseberlindner yeast Cyberlindenera jadinii 413.

[0051] In some embodiments, the culture temperature of the Saccharomyces jedinseberlindera 413 is 28°C-38°C.

[0052] In some embodiments, the culture temperature of the Saccharomyces jedinseberlindera 413 is 35°C-38°C.

[0053] In a second aspect, some embodiments of the present invention also provide the use of the yeast Jardine Seiberrin in fermenting corn to prepare protein feed.

[0054] In some embodiments, the corn syrup is liquefied, enzymatically hydrolyzed and saccharified, a nitrogen source is added, and a yeast solution of Saccharomyces cerevisiae is inoculated into the fermentation environment at 28°C-38°C with a ventilation volume of 1 m 3 / h-1.5m 3 / h, ferment for 28h-32h, after the fermentation is completed, separate the solid and liquid, and dry the solid to obtain protein feed; The Lindnerella jardiniseiber is Lindnerella jardiniseiber 413.

[0055] In some embodiments, the mass ratio of the bacterial liquid of Saccharomyces jadinseberlindnerii to the corn steep liquor in the fermentation environment is 5%.

[0056] In some embodiments, the specific method of the liquefaction treatment includes: adjusting the temperature and pH value of the corn slurry, and adding liquefaction enzyme treatment.

[0057] In some embodiments, the temperature of the corn steep liquor is adjusted to 91°C-97°C.

[0058] In some embodiments, the pH value is 5-6.

[0059] In some embodiments, the liquefaction enzyme comprises a liquefaction enzyme having an activity of 10,000-20,000 U / g.

[0060] In some embodiments, the mass concentration of the liquefaction enzyme in the corn steep liquor is 0.2%.

[0061] In some embodiments, an iodine test is performed on the corn slurry during the liquefaction process. When the substrate undergoing the liquefaction process reacts with the iodine solution and changes from blue to orange during the iodine test, the liquefaction process is completed.

[0062] In some embodiments, the specific method of reducing viscosity by enzymatic hydrolysis includes: adding enzymatic hydrolysate to the liquefied corn steep liquor, adjusting the pH, and heating in a water bath.

[0063] In some embodiments, the enzymatic hydrolysate includes one or more of cellulase and pectinase.

[0064] In some embodiments, the cellulase has an activity of 400,000-500,000 U / g.

[0065] In some embodiments, the pectinase has an activity of 10,000-20,000 U / g.

[0066] In some embodiments, the amount of the enzymatic hydrolysate added to the corn steep liquor after liquefaction is 0.2% by weight.

[0067] In some embodiments, the pH is adjusted to 4-5.

[0068] In some embodiments, the water bath is heated at a temperature of 50°C-55°C.

[0069] In some embodiments, the viscosity of the corn steep liquor during the enzymatic viscosity reduction process is measured, and when the viscosity is lower than 2000 mPa.s, the enzymatic viscosity reduction is completed.

[0070] In some embodiments, the specific method of the saccharification treatment includes: adding saccharifying enzyme to the corn syrup after enzymatic hydrolysis and viscosity reduction, adjusting the pH, and heating in a water bath.

[0071] In some embodiments, the amount of saccharifying enzyme added to the corn steep liquor after enzymatic hydrolysis and viscosity reduction is 0.6%.

[0072] In some embodiments, the saccharifying enzyme comprises a saccharifying enzyme having an activity of 40,000-50,000 U / g.

[0073] In some embodiments, the pH is adjusted to 4-5.

[0074] In some embodiments, the water bath is heated at a temperature of 55°C-60°C.

[0075] In some embodiments, the presence of dextrin in the corn slurry during the saccharification process is detected by an anhydrous ethanol method, and the saccharification process is terminated when no dextrin is present in the corn slurry during the saccharification process.

[0076] In some embodiments, the nitrogen source includes ammonium sulfate.

[0077] In some embodiments, the nitrogen source is added to the corn steep liquor in an amount of 1%-2% after saccharification.

[0078] In some embodiments, during fermentation, the gas introduced is air.

[0079] In some embodiments, the corn slurry is prepared by beating corn and water, and the mass ratio of corn to water is 1:1.25.

[0080] In some embodiments, the method for preparing the bacterial liquid of Saccharomyces jadiniensis 413 comprises: placing Saccharomyces jadiniensis 413 in a shake flask culture medium, and culturing at 28° C.-38° C. for 24 h-28 h.

[0081] In some embodiments, the formula of the shake flask culture medium is: yeast extract, glucose, magnesium sulfate, and dipotassium hydrogen phosphate; the mass concentration ratio of the yeast extract, glucose, magnesium sulfate, and dipotassium hydrogen phosphate in the shake flask culture medium is (8.5g / L-10.5g / L):(18g / L-21g / L):(1g / L-1.5g / L):(1.3g / L-1.5g / L).

[0082] In some embodiments, the drying temperature is 55°C-65°C.

[0083] The following is described in conjunction with specific embodiments: (1) Activation of the original strain After the original strain BNCC335990 was restored to room temperature, 1-2 rings of single colonies were picked from it and streaked on YPD medium to rejuvenate the strain. After the strain matured, 1-2 rings of vigorous single colonies were picked from it and inoculated into YPD liquid medium (commercially available) to activate the strain. Then it was placed in a shaker at 30°C, 165 rpm / min, and time for 16 hours. After that, the strain was serially diluted 10 times to make 10 -1 -10 -5 of bacterial suspension, take 10 -3 -10 -5 The bacterial suspensions were spread on culture dishes and cultured at a constant temperature of 30°C. The morphology and number of colonies were observed after culture, and the appropriate dilution multiple of the bacterial suspension was selected.

[0084] (2) Ultraviolet radiation mutagenesis The activated strain was inoculated into a new YPD liquid medium for shake flask culture. When the culture reached the logarithmic growth phase, 1 mL of the bacterial solution was taken and centrifuged at 6000 rpm for 10 min. The supernatant was removed and washed with sterile saline for 2-3 times. The suspension was centrifuged again at 6000 rpm for 8 min, the supernatant was removed and 1 mL of sterile saline was added to obtain a bacterial suspension. A 10-fold serial dilution method was used to prepare 10 -1 -10 -5 When the bacterial suspension reached the logarithmic growth phase, 1 mL of the bacterial suspension was spread on a plate and irradiated at a distance of 30 cm perpendicular to a UV lamp (15 W) for 0 s (control), 20 s, 40 s, 60 s, 80 s, 100 s, and 120 s, respectively. Finally, the plate was exposed to UV light for 10 -4 Under the condition of UV irradiation time of 120s, a plate with good colony distribution and a lethality rate that met the requirements (75%) was obtained, and the plate without UV treatment was used as the control.

[0085] (3) High temperature culture screening The strain obtained after UV mutagenesis was cultured at high temperature to obtain a high-protein accumulating strain that is resistant to high temperatures. The bacterial suspension after UV mutagenesis was diluted to 10 -4 , then the induced bacterial suspension was spread on YPD solid medium, inverted and cultured at 38°C for 48-72h, and several colonies with larger colonies and better colors were selected to obtain temperature-resistant primary screening strains. The screened strains were inoculated into 250mL liquid culture medium and cultured at 38°C and 165rpm until the logarithmic growth phase. The bacterial mass and protein content were measured respectively, and 4 prominent colonies were selected for measurement (i.e., A1-A4 in Table 1). The formula of the shake flask medium is: yeast extract, glucose, magnesium sulfate, dipotassium hydrogen phosphate, and water; the mass concentrations of the yeast extract, glucose, magnesium sulfate, and dipotassium hydrogen phosphate in the shake flask medium are 9.5g / L, 20.0g / L, 1.3g / L, and 1.4g / L, respectively.

[0086] Table 1 Test results of different prominent colonies

[0087] Finally, A3 was selected as the target strain. Under the culture condition of 38°C, its protein content was 45.2%, which was 48.2% higher than that of the control, and its biomass was 38.9 g / L, which was 34.6% higher than that of the control.

[0088] (4) Identification of bacterial species: Strain A3 was sequenced, and the 26 SrDNA gene D1 / D2 region sequence analysis method was used to extract the yeast sample DNA, PCR amplify, sequence and compare the gene sequence homology with the NCBI database. Finally, the strain was determined to be Saccharomyces jadinseberlindnerii ( Cyberlindnera jadinii ), and the strain was named Lindnerella jedinseiberi ( Cyberlindnera jadinii )413, deposited on November 11, 2024 at the Guangdong Microbial Culture Collection Center (GDMCC) at the Institute of Microbiology, Guangdong Academy of Sciences, 5th Floor, Building 59, No. 100, Xianlie Middle Road, Guangzhou, with the deposit number GDMCC No: 65456.

[0089] The mutagenized Jadinseberlindner yeast 413 was compared with the unmutated Jadinseberlindner yeast at different culture temperatures. Specifically, the strain was inoculated into a shake flask medium for culture. The shake flask culture conditions were: cultured at 30°C, 32°C, 34°C, 36°C, and 38°C, respectively, for 28 hours, and pH 5.5. The fermentation broth in the stable phase was obtained, and the number of viable bacteria in the fermentation broth was determined. The results are as follows: Figure 1 The formula of the shake flask medium is as follows: the concentrations of yeast extract, glucose, magnesium sulfate, and potassium dihydrogen phosphate in the shake flask medium are 10.5 g / L, 21 g / L, 1.5 g / L, and 1.5 g / L, and the balance is water.

[0090] Embodiment 1: Lindnerella jedinii 413 was inoculated into a shake flask medium for culture. The shake flask culture conditions were as follows: temperature 38°C, culture time 28h, pH 5.5, to obtain a fermentation broth in a stable phase. The shake flask medium formula was as follows: the concentrations of yeast extract, glucose, magnesium sulfate, and dipotassium hydrogen phosphate in the shake flask medium were 10.5g / L, 21g / L, 1.5g / L, and 1.5g / L, and the balance was water.

[0091] Fresh glutinous corn and water were mixed in a mass ratio of 1:1.25 to form a homogenate. The beating time was preferably 3-5 min. 300 g corn slurry was placed in a 500 mL conical flask. 3000 U / g liquefaction enzyme (Ningxia Heshibi Biotechnology Co., Ltd.) was added for liquefaction. The added mass was 0.2% of the mass of the corn slurry. The pH was adjusted to 5.9 and the liquefaction temperature was 95 °C. An iodine test was performed on the corn slurry during the liquefaction process. When the substrate of the liquefaction treatment reacted with the iodine solution during the iodine test and changed from blue to orange, the liquefaction treatment was completed. After the corn slurry after liquefaction treatment is cooled, 500,000 U / g of cellulase (Ningxia Heshibi Biotechnology Co., Ltd.) with an added mass of 0.1% of the corn slurry and 20,000 U / g of pectinase (Ningxia Heshibi Biotechnology Co., Ltd.) with an added mass of 0.1% of the corn slurry are added, the pH is adjusted to 4.5, and viscosity reduction treatment is carried out. The viscosity reduction temperature is 50°C. The viscosity of the corn slurry during the enzymatic viscosity reduction process is measured. When the viscosity is lower than 2000mpa.s, the enzymatic viscosity reduction is completed. After viscosity reduction, 50,000 U / g of saccharifying enzyme (Ningxia Heshibi Biotechnology Co., Ltd.) with an activeness is added for saccharification treatment. The pH is adjusted to 4.5, the added mass is 0.6% of the corn slurry, and the saccharification temperature is 60°C. The presence of dextrin in the corn slurry during the saccharification process is detected by the anhydrous ethanol method. When there is no dextrin in the corn slurry during the saccharification process, the saccharification process is completed.

[0092] The corn slurry after viscosity reduction and saccharification was inoculated with 5% of the corn slurry in the logarithmic phase of Saccharomyces cerevisiae 413 for fermentation. At the initial stage of fermentation, 2% of ammonium sulfate was added as a nitrogen source. The fermentation temperature was 38 °C, the fermentation time was 30 h, the fermentation pH was 5.5, and the air ventilation volume was 1 m 3 / h; after the fermentation is completed, the fermentation product is taken for centrifugal drying, the centrifugal speed is 8000rpm, the time is 6min, and the drying temperature is 60°C. The solid sample obtained is crushed into powder for the determination of protein content.

[0093] Embodiment 2: The preparation method of Example 2 is the same as that of Example 1, but the fermentation temperature in Example 2 is 30°C.

[0094] Embodiment 3: The preparation method of Example 3 is the same as that of Example 1, but the fermentation temperature in Example 3 is 32°C.

[0095] Embodiment 4: The preparation method of Example 4 is the same as that of Example 1, but the fermentation temperature in Example 4 is 34°C.

[0096] Embodiment 5: The preparation method of Example 5 is the same as that of Example 1, but the fermentation temperature in Example 5 is 36°C.

[0097] Comparative Example 1: The preparation method of Comparative Example 1 is the same as that of Example 1, but the strain used in Comparative Example 1 is before mutagenesis.

[0098] Comparative Example 2: The preparation method of Comparative Example 2 is the same as that of Example 2, but the strain used in Comparative Example 2 is before mutagenesis.

[0099] Comparative Example 3: The preparation method of Comparative Example 3 is the same as that of Example 3, but the strain used in Comparative Example 3 is before mutagenesis.

[0100] Comparative Example 4: The preparation method of Comparative Example 4 is the same as that of Example 4, but the strain used in Comparative Example 4 is before mutagenesis.

[0101] Comparative Example 5: The preparation method of Comparative Example 5 is the same as that of Example 5, but the strain used in Comparative Example 5 is before mutagenesis.

[0102] The protein in the protein feed obtained in the embodiment and the comparative example was tested: (1) Weigh 0.5g of the thoroughly mixed solid sample (equivalent to 30-40mg of nitrogen) to an accuracy of 0.001g and put it into the digestion tube. Then add 0.4g of copper sulfate, 6g of potassium sulfate and 20mL of sulfuric acid to the digestion furnace for digestion. When the temperature of the digestion furnace reaches 420℃, continue digestion for 1h. At this time, the liquid in the digestion tube is green and transparent. Take it out and cool it down. Add 50mL of water and perform the process of adding liquid, distilling, titrating and recording titration data on the Kjeldahl nitrogen analyzer. The calculation formula is as follows: F ; Where: X—protein content in the sample, in grams per hundred grams (g / 100g); V1—the volume of sulfuric acid or hydrochloric acid standard titrant consumed by the sample, in milliliters (mL); V2—the volume of sulfuric acid or hydrochloric acid standard titrant consumed by the sample blank, in milliliters (mL); C—concentration of sulfuric acid or hydrochloric acid standard titration solution, in moles per liter (mol / L); The mass of nitrogen equivalent to 0.0140-1mL of sulfuric acid or hydrochloric acid standard titration solution, in grams (g); m—mass of the sample, in grams (g); V3—the volume of digestive fluid absorbed, in milliliters (mL); F—nitrogen conversion factor to protein; 100—Conversion factor.

[0103] Results and Analysis: First, through Figure 1 It can be found that the Jadinseberlindner yeast 413 obtained by the present invention has good temperature resistance. In a fermentation environment where the fermentation temperature is greater than 34°C, the number of viable bacteria of the Jadinseberlindner yeast 413 of the present invention is significantly increased compared with the Jadinseberlindner yeast before induction, and the number of viable bacteria of the Jadinseberlindner yeast 413 at 30°C-38°C does not change much. It can be seen that the Jadinseberlindner yeast 413 of the present invention has good temperature resistance.

[0104] The use of Saccharomyces cerevisiae 413 in fermenting corn, such as Figure 2 As shown, it can be found that the Jardine Seiberlindner yeast 413 of the present invention has good temperature resistance activity, so it can better convert protein during the fermentation of corn. It can be seen that the Jardine Seiberlindner yeast 413 obtained by the present invention has good temperature resistance and can be extended to the fermentation environment with a fermentation temperature of 28°C-38°C for protein conversion.

[0105] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the technical solutions of the embodiments of the present invention.

Claims

1. A thermotolerant yeast, characterized in that The jadinseberlindner yeast is jadinseberlindner yeast 413; The Jadinseberlindner yeast 413 has been deposited in the Guangdong Provincial Microbial Culture Collection Center on November 11, 2024, with the deposit address being 5th Floor, Building 59, No. 100, Xianlie Middle Road, Guangzhou, with the deposit number being GDMCC NO: 65456, and the microbial classification is named Jadinseberlindner yeast Cyberlindenera jadinii 413.

2. The thermotolerant yeast Saccharomyces cerevisiae according to claim 1, characterized in that The culture temperature of the Lindnerella jardiniseiber 413 is 28°C-38°C.

3. The use of the yeast Jardine Seiberlindnerella according to any one of claims 1 to 2 in fermenting corn to prepare protein feed, characterized in that: The corn pulp was liquefied, enzymatically hydrolyzed to reduce viscosity, saccharified, and nitrogen source was added. The yeast solution of Saccharomyces cerevisiae was inoculated into the fermentation environment at 28℃-38℃ and the ventilation volume was 1m 3 / h-1.5m 3 / h, ferment for 28h-32h, after the fermentation is completed, separate the solid and liquid, and dry the solid to obtain protein feed; The Lindnerella jardiniseiber is Lindnerella jardiniseiber 413.

4. The use according to claim 3, characterized in that: The mass ratio of the bacterial liquid of the yeast Lindnerella jeddinghis to the corn steep liquor in the fermentation environment is 5%.

5. The use according to claim 3, characterized in that: The specific method of the liquefaction treatment includes: adjusting the temperature and pH value of the corn slurry, adding liquefaction enzyme treatment; Adjust the temperature of corn steep liquor to 91°C-97°C; The pH value is 5-6; The liquefaction enzyme includes a liquefaction enzyme with an activity of 10,000-20,000 U / g; The mass concentration of the liquefaction enzyme in the corn steep liquor is 0.2%.

6. The use according to claim 3, characterized in that: The specific method of reducing viscosity by enzymatic hydrolysis comprises: adding enzymatic hydrolysate to the liquefied corn steep liquor, adjusting pH, and heating in a water bath; The enzymolysate includes one or more of cellulase and pectinase; The amount of the enzymatic hydrolysate added to the corn steep liquor after liquefaction is 0.2% by weight; The pH is adjusted to 4-5; The water bath heating temperature is 50°C-55°C.

7. The use according to claim 3, characterized in that: The specific method of the saccharification treatment includes: adding saccharifying enzyme to the corn syrup after enzymatic hydrolysis and viscosity reduction, adjusting pH, and heating in a water bath; The added mass of the saccharifying enzyme in the corn steep liquor after enzymatic hydrolysis and viscosity reduction is 0.6%; The pH is adjusted to 4-5; The water bath heating temperature is 55°C-60°C.

8. The use according to claim 3, characterized in that: The nitrogen source includes ammonium sulfate; The nitrogen source is added in an amount of 1%-2% in the corn syrup after saccharification; During fermentation, the gas introduced is air; The corn slurry is prepared by: beating corn and water, wherein the mass ratio of corn to water is 1:1.25; The drying temperature is 55°C-65°C.

9. The use according to claim 3, characterized in that: The method for preparing the bacterial liquid of Saccharomyces jadiniensis berlindnerii comprises: placing Saccharomyces jadiniensis berlindnerii 413 in a shaking flask culture medium and culturing at 28° C.-38° C. for 24 h-28 h.

10. The use according to claim 9, characterized in that: The formula of the shake flask culture medium is: yeast extract, glucose, magnesium sulfate, and dipotassium hydrogen phosphate; the mass concentration ratio of the yeast extract, glucose, magnesium sulfate, and dipotassium hydrogen phosphate in the shake flask culture medium is (8.5g / L-10.5g / L):(18g / L-21g / L):(1g / L-1.5g / L):(1.3g / L-1.5g / L).

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