CN113832048B - Bacillus amyloliquefaciens for inhibiting soy sauce from forming white film and application thereof - Google Patents
Bacillus amyloliquefaciens for inhibiting soy sauce from forming white film and application thereof Download PDFInfo
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Abstract
The invention discloses bacillus amyloliquefaciens for inhibiting soy sauce from forming white films and application thereof, and belongs to the technical field of microorganisms. The invention obtains the strain bacillus amyloliquefaciens JDF-1 capable of inhibiting the formation of powdery pichia pastoris or white film produced by the pichia pastoris by separating and purifying the soy sauce mash microorganisms. In a fermentation system for simulating soy sauce, bacillus amyloliquefaciens JDF-1 is added to inhibit the generation of white films in the system. The bacillus amyloliquefaciens JDF-1 obtained by screening is suitable for the soy sauce fermentation production environment, and can well generate white films in soy sauce by adjusting the addition amount of the strain, so that the bacillus amyloliquefaciens JDF-1 has important significance for reducing the generation of the white films and improving the quality of the soy sauce in the soy sauce fermentation production.
Description
Technical Field
The invention relates to bacillus amyloliquefaciens for inhibiting soy sauce from forming white films and application thereof, and belongs to the technical field of microorganisms.
Background
Soy sauce production is a multi-strain fermentation process, and saccharomycetes are also important microorganisms for soy sauce brewing, and the strain plays an important role in the formation of soy sauce flavor. The yeast separated from the soy sauce mash comprises more than 30 species of zygosaccharomyces rouxii (Zygosaccharomyces rouxii), candida escitalopsis (Candida etchellsii), candida variabilis (Torulopsis versatilis), pichia farinose (Pichia farinosa) and the like. The zygosaccharomyces rouxii is a fermentation type yeast, mainly acts on the fermentation earlier stage, can convert glucose into ethanol, and can also produce various flavor substances such as esters, furanones and the like. The powdered pichia pastoris can produce alcohol in the soy sauce fermentation process, but the powdered pichia pastoris can form a white film on the surface of stationary soy sauce mash and consume part of amino acid, so that the quality of the soy sauce is reduced.
The advantageous characteristics of Pichia pastoris (Pichia farinosa) in the fermentation process, namely low pH resistance, low water activity resistance, high osmotic pressure resistance and short-time high temperature resistance, can lead to the problem that soy sauce sterilization cannot completely kill the Pichia pastoris, thereby increasing the problems of microorganism control and shelf life extension after fermentation. Currently, white films in soy sauce are mainly controlled in two aspects: 1. the fermentation process is improved. The production content of the white film is influenced by fermentation factors and fermentation technology, wherein the regulation and control of the oxygen content, temperature, salt concentration, pH and other factors can effectively reduce the production amount. The defects are that the workload is large, the time and the labor are consumed, and the efficiency is low due to too many influencing factors. 2. Microorganism means. Screening microorganisms which inhibit the growth of the white film-producing yeasts, which can block the production pathway from the source, is the most effective method at present, and modifying microorganisms so that the microorganisms can produce some metabolites to inhibit the growth of the white film-producing yeasts has the limitation that people worry about the safety of the strains and possibly have uncontrollable influence on the soy sauce flavor formation; therefore, screening strains capable of reducing the formation of white films during fermentation of soy sauce and using them for soy sauce production is currently the safest and effective method.
Disclosure of Invention
The invention provides bacillus amyloliquefaciens (Bacillus amyloliquefaciens) JDF-1 which is separated from soy sauce mash in the earlier stage of fermentation and can inhibit white film production of powdery pichia pastoris, and the strain is used for simulating soy sauce fermentation so as to solve the problem of soy sauce quality reduction caused by white film production in soy sauce fermentation.
The first object of the invention is to provide a bacillus amyloliquefaciens (Bacillus amyloliquefaciens) which is preserved in China Center for Type Culture Collection (CCTCC) with the preservation number of M2021212 in the year 3 and 8 of 2021.
The invention also provides a microbial preparation containing the bacillus amyloliquefaciens.
In one embodiment, the microbial preparation also contains other types of microorganisms useful for fermentation, including Aspergillus oryzae.
The invention also provides wheat starter, bran starter or other types of starter containing the bacillus amyloliquefaciens.
The invention also provides a method for culturing the bacillus amyloliquefaciens JDF-1, which comprises the steps of inoculating the bacillus amyloliquefaciens JDF-1 into a beef extract peptone culture medium, and carrying out static culture at 28-30 ℃.
The invention provides a method for inhibiting soy sauce white film production, which comprises the step of adding bacillus amyloliquefaciens JDF-1 into a soy sauce fermentation system for collaborative fermentation after the start of soy sauce fermentation.
The invention provides a Pichia farinosa (Pichia farinosa) CM-1 which has been preserved in China Center for Type Culture Collection (CCTCC) with the preservation number of M2021213 in 2021 and 3 months and 8 days.
The invention provides a method for culturing powdery pichia pastoris capable of producing white films in soy sauce fermentation, which comprises the steps of inoculating powdery pichia pastoris CM-1 into YPD culture medium and performing stationary culture at 30 ℃.
In one embodiment of the present invention, the concentration of the Bacillus amyloliquefaciens JDF-1 in the environment of 100g/L NaCl is at least 1.0X10 4 CFU/g。
In one embodiment, the pichia pastoris is fermented during soy sauce fermentation.
The invention provides a method for inhibiting white film-producing yeast, which is to add bacillus amyloliquefaciens JDF-1 for fermentation in the soy sauce fermentation process.
In one embodiment, the Bacillus amyloliquefaciens JDF-1 is added to the fermentation system in an amount of 1.0X10% of the final concentration of the system 3 ～1.0×10 5 CFU/g。
The invention provides a micro-starter, which is prepared by the following steps: inoculating 200-600 mu L of bacillus amyloliquefaciens JDF-1 into 10-30 mL of beef extract peptone liquid medium, activating for 2-3 generations at 30 ℃, and waiting for the bacillus amyloliquefaciens JDF-1 to reach 1.0x10 5 Centrifuging at 6000-1000 rpm for 15min when the viable count is above CFU/mL, removing supernatant, sequentially adding buffer solution and cryoprotectant in sterile environment, and keeping cell concentration at not lower than 1.0X10 5 And (3) at the time of CFU/mL, performing vacuum freeze drying treatment to obtain the starter.
In one embodiment, the buffer solution is 0.1-0.3M phosphate buffer solution with pH value of 6-7 and/or 0.5-1% physiological saline and/or double distilled water, and the cryoprotectant is 10-20% glycerol and/or 8-12% skim milk powder and/or 8-12% trehalose.
The invention provides a method for preparing soy sauce without white film, which is to add the bacillus amyloliquefaciens JDF-1 or the starter in the soy sauce fermentation process for fermentation.
In one embodiment, the method comprises the steps of: bean pulp flour was mixed with 1: mixing at a ratio of 1, inoculating 2%yeast extract, making yeast at 30deg.C for 48 hr, mixing 20% saline water with yeast at a ratio of 1:1.8 (W/V) mixing, adding bacillus amyloliquefaciens JDF-1 or the starter for fermentation, wherein the fermentation temperature is 25 ℃ initially, and the temperature is 1 ℃ to 30 ℃ every two days until the fermentation is finished, and fermenting for 30 days.
In one embodiment, the bacillus amyloliquefaciens, or the starter, has a bacterial concentration of 1.0X10 in the fermentation system 3 ～1.0×10 5 CFU/g。
The invention also provides application of the bacillus amyloliquefaciens JDF-1 or the method for inhibiting white film production, or the starter, or the method for preparing soy sauce without white film production in preparing fermented food.
In one embodiment of the present invention, the alcoholic beverage comprises beer, red wine, fruit wine; the beverage includes a fermented juice.
The beneficial effects are that:
the invention obtains the strain bacillus amyloliquefaciens JDF-1 capable of inhibiting the formation of powdery pichia pastoris or white film produced by the pichia pastoris by separating and purifying the soy sauce mash microorganisms. In a fermentation system for simulating soy sauce, bacillus amyloliquefaciens JDF-1 is added to inhibit the generation of white films in the system. The bacillus amyloliquefaciens JDF-1 obtained by screening is suitable for the soy sauce fermentation production environment, and can well generate white films in soy sauce by adjusting the addition amount of the strain, so that the bacillus amyloliquefaciens JDF-1 has important significance for reducing the generation of the white films and improving the quality of the soy sauce in the soy sauce fermentation production.
Preservation of biological materials
Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) JDF-1 was designated by the classification: bacillus amyloliquefaciens (Bacillus amyloliquefaciens) JDF-1 was deposited at China Center for Type Culture Collection (CCTCC) No. M2021212 at the 3 rd month and 8 th year of 2021 with the deposit address of China, wuhan university.
A strain of Pichia farinosa (Pichia farinosa) CM-1, classified and named Pichia farinosa (Pichia farinosa) CM-1, was deposited in China center for type culture Collection, with a deposit number of CCTCC NO: M2021213, and a deposit address of China, wuhan, university of Wuhan.
Drawings
FIG. 1 film production of Pichia pastoris powder under soy sauce and culture medium conditions.
FIG. 2 shows the white film and growth of Pichia farinosa under different conditions; wherein, a is the influence of different salt concentrations on the growth of CM-1 b is the influence of different temperatures on the growth of CM-1 c is the influence of different pH values on the growth of CM-1.
FIG. 3 effects of different environmental factors on Bacillus amyloliquefaciens growth; wherein, a is the influence of different salt concentrations on the growth of the JDF-1 b is the influence of different temperatures on the growth of the JDF-1 c is the influence of different pH values on the growth of the JDF-1.
FIG. 4 inhibition of yeast by different moromi-derived microorganisms; wherein, a, the influence of microorganisms from different soy sauce mash sources on the growth of powdery pichia pastoris. And b, the effect of microorganisms from different soy sauce mash sources on the growth of candida.
FIG. 5 Bacillus amyloliquefaciens inhibits white film production by powdered Pichia pastoris.
FIG. 6 effect of Bacillus amyloliquefaciens on soy fermentation; wherein a, the influence of the addition of bacillus amyloliquefaciens JDF-1 on reducing sugar; b, adding the influence of bacillus amyloliquefaciens JDF-1 on amino acid nitrogen; and c, adding the influence of bacillus amyloliquefaciens JDF-1 on total acid.
Detailed Description
Beef extract peptone medium: 10g/L peptone, 3g/L beef extract and 5g/L sodium chloride.
YPD medium: 20g/L peptone, 10g/L yeast extract and 20g/L glucose. Simulated fermentation medium: the soybean meal and wheat flour used for producing soy sauce are mixed according to the weight ratio of 1: mixing at a ratio of 1, adding 4 times of water (w/v) and high-temperature amylase (50U/kg), steaming and gelatinizing for 1h, rapidly cooling to 60deg.C, adding saccharifying enzyme (120U/kg), and saccharifying at 60deg.C for 2h to obtain simulated fermentation medium.
The soy sauce referred to in the present application is, unless otherwise specified, a liquid soy sauce obtained by filtering moromi. The soy sauce mash is a semi-finished product in the fermentation process after adding salt water in the yeast.
Example 1: white film produced by powdered pichia pastoris in soy sauce and culture medium
Taking 250mL of fermented soy sauce and 250mL of YPD medium, respectively 1.0X10 5 The CFU/mL added with powdered Pichia pastoris CM-1, without any strain added, was used as a negative control. And (3) standing and fermenting for 3d at 30 ℃ compared with a control without adding powdery pichia pastoris. The addition of powdered pichia pastoris to the medium and soy sauce produced white films (see figure 1).
Example 2: white film produced by powdery pichia pastoris under different conditions and growth conditions
(1) Growth and film production conditions of powdery pichia pastoris under different sodium chloride concentrations
Inoculating the strain into YPD liquid, standing at 30deg.C for 3d, collecting 1mL of bacterial liquid 10000 r.min -1 Centrifuging for 10min. Washing twice with 0.8% (w: v) sterile physiological saline, re-suspending with 1mL sterile water, adding the bacterial liquid into YPD medium containing 0%,5%,10%,15%, and 20% NaCl at 10% (w: v) inoculum size, respectively, standing at 30deg.C, and culturing at initial and final viable count of C 0 And C 1 Expressed in CFU.mL -1 Relative growth rate= (lg C 1 -lg C 0 )/24. The result shows that the relative growth rate gradually decreases along with the increase of the salt concentration, but the powdery pichia pastoris CM-1 can also maintain 45% relative growth rate under the 18% salt concentration, and the powdery pichia pastoris is proved to be a strain of salt-tolerant bacteria.
(2) Growth and film production conditions of powdery pichia pastoris under low temperature condition
The activated bacterial cells were inoculated into YPD medium at an inoculum size of 10% (w: v), cultured at 15℃and 20℃for 3 days, sampled, spread in a gradient manner, and viable cell count was performed. Viable count at the start and end of culture was C 0 And C 1 Expressed in CFU.mL -1 Relative growth rate= (lg C 1 -lg C 0 )/24。
The result shows that the powdered pichia pastoris has different relative growth rates at different temperatures, the highest relative growth rate can reach 82% at 30 ℃, and the 41% relative growth rate can be maintained at 15 ℃, which proves that the powdered pichia pastoris CM-1 can grow at low temperature.
(3) Growth and film production conditions of powdery pichia pastoris under different pH conditions
Adding the activated bacterial liquid into YPD culture medium with pH of 5.0, 5.5, 6.0, 6.5, 7.0 according to 10% (w: v) inoculum size, standing at 30deg.C for culturing, and collecting viable cell count at the beginning and end of culture as C 0 And C 1 Expressed in CFU.mL -1 Relative growth rate= (lg C 1 -lg C 0 )/24。
The result shows that the relative growth rate of the powdery pichia pastoris is up to 84% when the pH value is 5.5, and can keep 45% when the pH value is 5.0, thus proving that the powdery pichia pastoris CM-1 can grow and reproduce rapidly in a meta-acid environment.
Example 3: screening for inhibiting bacillus amyloliquefaciens produced by powdery pichia pastoris
Taking 5g of soy sauce mash in 100mL of beef extract peptone liquid medium filled with glass beads, shaking uniformly, and standing and culturing for 1-3d at 37 ℃; centrifuging the supernatant at 12000rpm at 4deg.C for 5 min; gradient dilution of supernatant 10 -2 、10 -3 、10 -4 、10 -5 、10 -6 Sucking 0.1mL of each gradient bacteria liquid, respectively coating the gradient bacteria liquid on a beef extract peptone culture medium, and culturing for 1-3d at 37 ℃; picking colonies with different forms, respectively marking on a beef extract peptone solid culture medium, and then standing and culturing at 30 ℃ for 1-3d until the colonies grow; single colonies of each strain were obtained after streaking and separation 3 times according to the above procedure. Single colony liquid culture is carried out, 1% of inoculation amount is inoculated into a culture medium containing powdery pichia pastoris, standing culture is carried out for 3d, and film production condition is observed.
Separating bacillus amyloliquefaciens capable of inhibiting white film production of powdery pichia pastoris in a culture medium. Bacterial genomic DNA was extracted from the bacterial solution using a Tiangen company bacterial genomic DNA extraction kit. PCR amplification was performed using the universal primers 27F and 1492R of the bacterial 18S rRNA gene, and the PCR product was sent to the Sesamum indicum sequencing company for sequencing. And submitting the 16S rRNA gene sequence obtained by sequencing to GenBank for BLAST comparison, and identifying the bacterial species.
27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-GGTTACCTTGTTA CGACTT-3')
PCR reaction System (50. Mu.L): 50 mg.L -1 Template 2. Mu.L, 10. Mu. Mol.L -1 1. Mu.L of each of the upstream and downstream primers, pfu PCR Master Mix. Mu.L of each of the upstream and downstream primers, and 21. Mu.L of double distilled water. Reaction conditions: 94 ℃ for 3min;94℃1min,55℃1min,72℃2min,30 cycles; and at 72℃for 10min. The PCR product was sent to Tien without tin for sequencing, and the sequencing results were aligned in the Genbank database of NCBI.
Table 1 results of screening the obtained strains
Example 4: inhibition ability of Bacillus amyloliquefaciens to different strains
(1) The mutual bacteriostasis of each microorganism is detected by adopting an oxford cup method.
The strain culture method comprises the following steps: yeast (Pichia farinosa, C.Parapsilosis) was cultured in YPD medium at 30deg.C 220 r.min -1 Culturing lactic acid bacteria (W.parametesenteroides, P.acidophilic, T.halophilius) at 37deg.C in MRS medium, and culturing bacillus at 37deg.C in LB medium for 1-2d.
Culturing Bacillus amyloliquefaciens in MRS culture medium at 37deg.C, and concentrating to 10 7 CFU·mL -1 Is used for fermenting a proper amount of fermentation liquor in 12000 r.min -1 Centrifuging at 4deg.C for 5min, collecting supernatant, and filtering with 0.22 μm filter membrane.
(2) Will be cultivated to logarithmic phase (bacterial concentration 10) 7 CFU·mL -1 ) Pichia farinosa and C.Parapsilosis are mixed with soft agar medium at a ratio of 1:50 (v: v), and the mixture is poured onto the lower medium with oxford cup. Taking out oxford cups by forceps after the soft agar culture medium is solidified, adding 100 mu L of bacillus amyloliquefaciens B.amyloliquefaciens JDF-1 supernatant prepared in the step (1) into corresponding holes, taking the holes added with sterile water as a control, placing the plates in a proper constant temperature incubator for culturing for 20 hours, and examining whether the strains have inhibition effect on each indicator bacterium. Each indicator bacterium3 parallels were made. Wherein the indicator bacteria are various yeasts, lactobacillus, etc. derived from soy sauce. The result shows that the bacillus amyloliquefaciens JDF-1 has remarkable inhibition effect on the P.farinosa strain, and the inhibition capability is superior to that of another bacillus amyloliquefaciens JY06, wherein the inhibition zone on the powdery pichia pastoris of the white film is maximum and reaches 2.12cm.
TABLE 2 inhibition of membrane-producing yeasts by different strains
Example 5: culturing Bacillus amyloliquefaciens under different environments
(1) Culturing bacillus amyloliquefaciens JDF-1 in high concentration sodium chloride environment
Inoculating the strain into MRS liquid, standing at 37deg.C for 24 hr, collecting 1mL of bacterial liquid 10000 r.min -1 Centrifuging for 10min. Washing twice with 0.8% (w: v) sterile physiological saline, re-suspending with 1mL sterile water, adding the bacterial solutions to MRS culture medium containing 0%,5%,10%,15%, and 20% NaCl at 10% (w: v) respectively, standing at 37deg.C, and culturing at initial and final viable count of C 0 And C 1 Expressed in CFU.mL -1 Relative growth rate= (lg C 1 -lg C 0 )/24. The result shows that the relative growth rate gradually decreases along with the increase of the salt concentration, but the bacillus amyloliquefaciens JDF-1 can also maintain 57 percent of relative growth rate at 15 percent of salt concentration, and the bacillus amyloliquefaciens JDF-1 is proved to be a salt-tolerant bacterium.
(2) Culturing bacillus amyloliquefaciens JDF-1 under low temperature condition
The activated bacterial liquid was added to MRS medium at an inoculum size of 10% (w: v), cultured at 15℃and 20℃for 24 hours, sampled, spread in a gradient manner, and viable bacteria counted. Viable count at the start and end of culture was C 0 And C 1 Expressed in CFU.mL -1 Relative growth rate= (lg C 1 -lg C 0 )/24。
The result shows that the bacillus amyloliquefaciens has different relative growth rates at different temperatures, the highest relative growth rate can reach 82% at 25 ℃, and the relative growth rate can be kept at 41% at 15 ℃, which proves that the bacillus amyloliquefaciens JDF-1 can grow at low temperature.
(3) Culturing Bacillus amyloliquefaciens JDF-1 under different pH conditions
Adding the activated bacterial liquid into MRS culture medium with pH of 5.0, 5.5, 6.0, 6.5, 7.0 according to 10% (w: v) inoculum size, standing at 37deg.C for culturing, and collecting viable cell count at the beginning and end of culture as C 0 And C 1 Expressed in CFU.mL -1 Relative growth rate= (lg C 1 -lg C 0 )/24。
The result shows that the relative growth rate of the bacillus amyloliquefaciens JDF-1 is up to 84 percent at the pH value of 7.0, and can keep 45 percent at the pH value of 5.0, thus proving that the bacillus amyloliquefaciens JDF-1 can survive in the environment of acid bias.
Example 6: bacillus amyloliquefaciens for inhibiting growth of powdery pichia pastoris and producing white film
In 250mL YPD medium, 1.0X10 5 CFU/mL is added with powdered Pichia pastoris and candida glabrata, and the ratio is 1.0X10 respectively 5 CFU/mL was added with B.amyloquefaciens JDF-1, W.parametesenteroides LCW-28, P.acidophilic B1419, T.halophilius R44, with Candida glabrata as positive control, without any strain added as negative control. And (3) standing and fermenting for 4d at 30 ℃ (figure 4).
In YPD medium and soy sauce, 1.0X10 respectively 5 The CFU/mL is added with bacillus amyloliquefaciens JDF-1 and powdered Pichia pastoris, only the powdered Pichia pastoris is added as a positive control, and no strain is added as a negative control. And (3) standing and culturing for 3d at 30 ℃.
Bacillus amyloliquefaciens JDF-1 can significantly inhibit the growth of Pichia pastoris CM-1, but cannot inhibit the growth of candida glabrata (no significant difference), and the addition of Bacillus amyloliquefaciens JDF-1 can prevent the generation of white films in the system (see FIG. 5).
Example 7: white membrane inhibiting bacillus amyloliquefaciens in simulated fermentation
To simulate hairThe fermentation medium is an examination system for soy sauce mash fermentation. In order to eliminate the interference of other microorganisms in the soy sauce mash on experimental results, a simulated fermentation medium is taken, sterilized for 20min at 121 ℃ and then uniformly mixed, and the ratio of 1:1.8 (w/v) saline with a concentration of 200g/L NaCl was added in such a ratio that the final concentration of the salt was 18%. The final concentration of each of the above-mentioned fermented soy sauce is 1.0X10% before the start of fermentation 4 CFU/g of powdered Pichia pastoris and bacillus amyloliquefaciens JDF-1, and the effect of inhibiting white film production by the bacillus amyloliquefaciens JDF-1 is verified. The fermentation temperature was initially 25℃and was increased every two days at 1℃to 30℃until the end of the fermentation, and during the fermentation for 30 days, the addition of Bacillus amyloliquefaciens JDF-1 reduced the production of white films in the system compared to the control without the addition of Bacillus amyloliquefaciens, and the amino acid nitrogen of soy sauce reached 1.24g/100mL without significant difference from the control (without any bacteria normally added) 1.2g/100mL, the reducing sugar content was 17.05g/L without significant difference from the control (17.22 g/L), and the total acid content was 5.4g/L without significant difference from the control (5.6 g/L) (FIG. 6).
Example 8: preparation of soy sauce without white film
Bean pulp flour was mixed with 1: mixing at a ratio of 1, inoculating 2%yeast extract, making yeast at 30deg.C for 48 hr, mixing 20% saline water with yeast at a ratio of 1:1.8 (W/V) mixing and adding the mixture to a final concentration of 1.0X10 3 ～1.0×10 5 CFU/g bacillus amyloliquefaciens JDF-1 or a starter containing the bacillus amyloliquefaciens JDF-1 is fermented, the fermentation temperature is initially 25 ℃, and the fermentation temperature is increased to 30 ℃ every two days at 1 ℃ until the fermentation is finished, and the fermentation is carried out for 30 days. The results showed that no white film was seen on the surface of the fermented soy sauce.
Comparative example 1:
the specific embodiment is the same as in example 6, except that Bacillus amyloliquefaciens JDF-1 is replaced by Bacillus amyloliquefaciens JY06 screened in example 1 or Weissella paramesenteroides LCW-28,Tetragenococcus halophilus R44 or Pediococcus acidilactici B1419 deposited in the laboratory, and the results show that W.parametes LCW-28, P.acidophilus B1419 and T.halophilius R44 can not inhibit the growth of powdery pichia pastoris nor candida glabra, and have no obvious effect on the reduction of white films of soy sauce systems.
While the invention has been described with reference to the preferred embodiments, it is not limited thereto, and various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
SEQUENCE LISTING
<110> Zhuhai Tianhe food Co., ltd
Jiangnan University
<120> Bacillus amyloliquefaciens for inhibiting soy sauce from forming white film and application thereof
<130> BAA210708A
<160> 2
<170> PatentIn version 3.3
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<211> 223
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<213> Bacillus amyloliquefaciens
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<213> Pichia farinosa
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Claims (9)
1. Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) JDF-1 has been deposited in China Center for Type Culture Collection (CCTCC) with the accession number of M2021212 at day 3 and 8 of 2021.
2. A composition comprising bacillus amyloliquefaciens JDF-1 of claim 1, wherein the composition is a wheat starter, a bran starter, or other types of starter.
3. A starter culture comprising the Bacillus amyloliquefaciens JDF-1 of claim 1.
4. A process for preparing the fermenting agent according to claim 3, wherein the Bacillus amyloliquefaciens JDF-1 is cultured, the cells are centrifuged and collected, a buffer solution and/or a protective agent are added to obtain a cell heavy suspension, and the heavy suspension is subjected to freeze drying to obtain the fermenting agent.
5. A method for inhibiting white film formation in soy sauce, which comprises adding Bacillus amyloliquefaciens JDF-1 according to claim 1 to soy sauce for co-fermentation before the start of fermentation of soy sauce.
6. The method according to claim 5, wherein the Bacillus amyloliquefaciens JDF-1 is added to the fermentation system in an amount of 1.0X10 3 ～1.0×10 5 CFU/g。
7. A method for producing soy sauce, characterized in that after koji making, koji is mixed with brine, the bacillus amyloliquefaciens JDF-1 of claim 1 or the composition of claim 2 is added to the mixed system, and fermented at a certain temperature.
8. The method according to claim 7, wherein the fermentation is carried out at an initial temperature of 25 ℃ to 30 ℃ at a rate of 1 ℃ every two days, and is maintained at 30 ℃ for a certain period of time.
9. Use of bacillus amyloliquefaciens JDF-1 according to claim 1, or a composition according to claim 2, or a starter culture according to claim 3, in soy sauce production.
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