CN115404173A - Aspergillus oryzae ZA216 and application thereof - Google Patents
Aspergillus oryzae ZA216 and application thereof Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/50—Soya sauce
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/60—Salad dressings; Mayonnaise; Ketchup
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/065—Microorganisms
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/66—Aspergillus
- C12R2001/69—Aspergillus oryzae
Abstract
The invention relates to the technical field of microbial fermentation, in particular to an aspergillus oryzae ZA216 and application thereof. The Aspergillus oryzae ZA216 is preserved in Guangdong province microorganism culture collection center at 2021, 5 months and 18 days, and the preservation number is GDMCC NO:61673, deposit address: guangzhou city, first furious Zhonglu No. 100 large building No. 59. The aspergillus oryzae ZA216 disclosed by the invention has long hypha, is good in water absorption, facilitates the enzyme in the yeast to diffuse and act on particles of the sauce mash, better decomposes the nutritional ingredients in the raw materials and improves the utilization rate of the raw materials. On the other hand, the soy sauce obtained by fermentation has high protease activity, high amino acid nitrogen and total nitrogen of the soy sauce, and the aroma and the taste of the soy sauce obtained by fermentation are obviously superior to those of the Shanghai brewing 3.042 strain soy sauce, wherein the soy sauce has rich and abundant aroma, prominent delicate flavor and mellow and long-lasting taste.
Description
Technical Field
The invention relates to the technical field of microbial fermentation, in particular to an aspergillus oryzae ZA216 and application thereof.
Background
Soy sauce is one of the oldest seasonings in the world and has been in china for over 2500 years. Soy sauce and products thereof have become a necessary seasoning for the life of not only traditional consumer countries of asia such as china, japan and korea due to their good taste and flavor, nutritional ingredients and biofunctional activity, but also has been gaining acceptance and favor from consumers all over the world. The preparation process of the soy sauce can be divided into three stages, namely, starter propagation of raw materials, fermentation of soy sauce mash and separation of the soy sauce. Koji making is a very important part of soy sauce production, and the quality of soy sauce koji plays an extremely important role in soy sauce production. The protease activity is one of key indexes for judging the quality of soy sauce koji, and has obvious influence on the soy sauce oil yield quality and the improvement of the raw material utilization rate.
At present, the price of raw materials rises seriously in the world, great challenges are brought to the seasoning industry, the requirement for improving the utilization rate of the raw materials is more and more clear, the main means for achieving the aim is to improve the protease activity of the soy sauce koji material, and the protease activity mainly depends on koji-making strains. Therefore, the acquisition of a high-yield protease strain is extremely important.
The process by which filamentous fungi secrete proteins by exocytosis usually occurs at the apical or sub-apical end of the hyphae, i.e., apical secretion. A small portion of protein secretion also occurs in other parts of the cell, such as the septum. Hyphae deposit new plasma membrane and cell wall components on the hyphal tip through exocytosis of secretory vesicles to realize tip elongation. Both longer apical-extending hyphae and highly branched hyphae are often associated with higher protein secretion capacity. There is a lot of evidence that the hyphal morphology of fungi can influence the production of extracellular proteins, and strains with altered part of the hyphal morphology show higher enzyme production capacity. Optimizing the mycelium morphology of filamentous fungi has important significance for improving protein synthesis and secretion. In addition, longer hypha can be more beneficial to extending into the raw materials such as soybean and wheat in the fermentation process of soy sauce starter propagation, so that the raw materials are better cracked and digested, and the utilization rate of the raw materials is improved.
The polar growth and hyphal morphology of filamentous fungi can be regulated by adjusting culture conditions. In the traditional fermentation industry, the mycelium morphology is generally controlled by setting fermentation process parameters such as seed amount, rotating speed, temperature, pH and the like. The method is relatively suitable for liquid fermentation systems, and is difficult to realize in solid fermentation systems such as soy sauce starter propagation and the like. The patent recombinant expression vector for changing the hypha form of trichoderma reesei and the application thereof changes the hypha form of trichoderma reesei by carrying out genetic engineering modification on the trichoderma reesei, is beneficial to forming more hypha end points, further improves the protein secretion capacity of the trichoderma reesei, and simultaneously influences the viscosity and dissolved oxygen of the trichoderma reesei fermentation liquor in the fermentation process, further influences the cellulase production capacity of the trichoderma reesei fermentation liquor. However, the patent uses genetic engineering means and is not suitable for being applied to the food industry.
In the patent of Trametes versicolor laccase fermentation process, ultrasonic treatment is carried out on thalli in a shake flask by using high-intensity ultrasonic waves, so that hyperbranched hypha forms and corm tops of hyphae of the Trametes versicolor laccase are generated to increase the area of secretion sites, and the permeability of cell membranes is improved to accelerate the secretion of laccase. The method adopts ultrasonic means for treatment, is difficult to expand the application to production, and increases the fermentation process and cost.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides an Aspergillus oryzae ZA216 and application thereof, and the invention screens out the mycelium length of the Aspergillus oryzae ZA216, on one hand, the water absorption is good, the enzyme in the yeast is convenient to diffuse and act on the particles of the soy sauce mash, the nutrient components in the raw materials are better decomposed, and the utilization rate of the raw materials is improved. On the other hand, the soy sauce koji protease obtained by fermentation has high activity and high amino acid nitrogen and total nitrogen of the soy sauce.
In order to realize the purpose, the invention adopts the technical scheme that:
in a first aspect, the present invention provides an Aspergillus oryzae (Aspergillus oryzae) ZA216 deposited at the southern east China collection of microorganisms at 18/5/2021 under the accession number GDMCC NO:61673, the preservation address is No. 59 building of Jieli Zhonglu 100 Dazhong Yao, guangzhou city.
The colony characteristics of Aspergillus oryzae (Aspergillus oryzae) ZA216 screened by the present invention are as follows: on the bean juice culture medium, hyphae grow vigorously, and aerial hyphae are white and fluffy velvet at the initial culture stage; along with the prolonging of the culture time, aerial hyphae grow radially from the center of the bacterial colony to the periphery, and the diameter of the bacterial colony is increased; the growth height of the aerial hyphae is gradually reduced from the center of the bacterial colony to the edge of the bacterial colony, the conidia are changed into yellow green from yellow, the mature spores are yellow green, and the back of the bacterial colony is tawny and has folds.
The Aspergillus oryzae ZA216 of the present invention is a strain obtained by breeding by mutation using Huniang 3.042 as a starting strain.
Preferably, the method for screening Aspergillus oryzae (Aspergillus oryzae) ZA216 comprises the steps of:
aspergillus oryzae (Aspergillus oryzae) ZA216 is prepared by using Hui brewing 3.042 as starting strain, subjecting the starting strain to mutagenesis by known mutagenesis method (such as ARTP mutagenesis), and coating on soybean protein plate to obtain Aspergillus oryzae variant strain; the selected Aspergillus oryzae strain variegated koji was cultured by the conventional culture method of Shanghai brewing 3.042 strain, and the growth of Aspergillus oryzae was observed. Selecting Aspergillus oryzae variant strain with high growth speed, long hypha and rich spore, performing triangular flask fermentation, and transplanting on bean juice slant culture medium for strain preservation. And (3) making the selected long hypha and high protease viable strains by using soybean/defatted soybean and wheat as raw materials by a conventional method, and carrying out a soy sauce fermentation pilot test. A strain which has high growth speed, long hyphae and high protease activity of the soy sauce koji obtained by fermentation in a small test is screened out, namely the Aspergillus oryzae (Aspergillus oryzae) ZA216.
Through genetic stability experiments, the amino acid nitrogen and total nitrogen contents of the fermented soy sauce of the screened Aspergillus oryzae (Aspergillus oryzae) ZA216 pass through generations 1, 5 and 10, and the amino acid nitrogen and total nitrogen contents are approximately kept unchanged, which shows that the Aspergillus oryzae (Aspergillus oryzae) ZA216 has good genetic stability.
In a second aspect, the present invention provides the use of Aspergillus oryzae ZA216 as described above for the fermentation of flavourings.
When the Aspergillus oryzae (Aspergillus oryzae) ZA216 fermentation seasoning is adopted to terminate fermentation, the amino acid nitrogen and the total nitrogen are improved by 6 percent and 2 percent compared with the original strain Shanghai brewing 3.042. The prepared seasoning is remarkably superior to 3.042 strain seasoning in the aspects of aroma and mouthfeel, wherein the aroma is rich and abundant, the delicate flavor is outstanding, and the mouthfeel is mellow and long-lasting.
As a preferred embodiment of the use according to the invention, the seasoning is soy sauce or sauce.
In a third aspect, the present invention provides the use of Aspergillus oryzae ZA216 as defined above in the preparation of koji.
As a preferred embodiment of the application of the present invention, the koji obtained by koji-making is used for preparing soy sauce or sauce.
In a fourth aspect, the present invention provides a koji prepared by inoculating Aspergillus oryzae (Aspergillus oryzae) ZA216 described above into a koji material prepared from soybean/defatted soybean, wheat.
The protease activity of the koji fermented by inoculating Aspergillus oryzae (Aspergillus oryzae) ZA216 of the invention is higher than that of the koji fermented by Humian 3.042 strain.
In a fifth aspect, the present invention provides the use of Aspergillus oryzae ZA216 as described above for improving the raw material utilization in soy sauce or sauce fermentation.
The mycelium of the Aspergillus oryzae ZA216 screened by the invention has good water absorption, facilitates the enzyme in the koji to diffuse and act on the granules of the soy sauce mash, better decomposes the nutritional ingredients in the raw materials and improves the utilization rate of the raw materials. On the other hand, the soy sauce koji protease obtained by fermentation has high activity, and the amino acid nitrogen and total nitrogen of the soy sauce are high.
In a sixth aspect, the present invention provides a method for producing the above soy sauce or paste, which comprises using Aspergillus oryzae ZA216 as a fermenting microorganism.
The Aspergillus oryzae ZA216 screened by the method is used as zymocyte, the growth speed is high, the culture is easy, and then seasonings such as soy sauce or sauce are obtained through fermentation, the obtained soy sauce or sauce is obviously superior to 3.042 strain seasonings in fragrance and mouthfeel, wherein the fragrance is strong and rich, the delicate flavor is prominent, the taste is mellow and long, and the quality is better.
As a preferred embodiment of the method for producing a soy sauce or a sauce of the present invention, the seasoning is a soy sauce or a sauce.
In a seventh aspect, the present invention provides a soy sauce or a paste which is produced by using the above Aspergillus oryzae ZA216 as a fermenting fungus.
The Aspergillus oryzae ZA216 of the invention has the advantages of high growth speed, easy culture, prominent flavor of fermented products, better quality, very large production and application value in soy sauce brewing and wide application prospect.
Compared with the prior art, the invention has the following beneficial effects:
the invention provides an Aspergillus oryzae ZA216 and application thereof, wherein the Aspergillus oryzae ZA216 has long hypha, so that the water absorption is good, enzyme in a koji material can be conveniently diffused and acts on particles of soy sauce mash, nutritional ingredients in raw materials can be better decomposed, and the utilization rate of the raw materials is improved. On the other hand, the soy sauce koji protease obtained by fermentation has high activity, high amino acid nitrogen and total nitrogen of the soy sauce, and the flavor and the taste of the soy sauce obtained by fermentation are obviously superior to those of the Shanghai brewing 3.042 strain soy sauce, wherein the soy sauce has strong and rich flavor, prominent delicate flavor and mellow and long-lasting taste.
Drawings
FIG. 1 is a diagram showing the hyphal length patterns of Humian 3.042 strain and Aspergillus oryzae ZA216 (the left is Humian 3.042 strain; the right is Aspergillus oryzae ZA 216);
FIG. 2 is a schematic representation of the form of Aspergillus oryzae ZA 216;
FIG. 3 is a graph showing the results of sensory analysis of crude oil fermented by the strain Hu Niang 3.042 and Aspergillus oryzae (Aspergillus oryzae) ZA216.
Detailed Description
To better illustrate the objects, aspects and advantages of the present invention, the present invention will be further described with reference to the accompanying drawings and specific examples.
In the following examples, the experimental methods used were all conventional methods unless otherwise specified, and the materials, reagents and the like used were commercially available unless otherwise specified.
Example 1 mutagenesis of Aspergillus oryzae strains
Spores of the Huniang 3.042 strain which is mature in culture are made into spore suspension, and ARTP mutagenesis is carried out on the spore suspension. Diluting spore suspension subjected to ARTP mutagenesis to 10 times by using sterile physiological saline according to a 10-fold gradient dilution method -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 And (5) concentration, and preparing a spore diluent. And applied to a soybean protein culture medium (the preparation method comprises 1g of soybean protein, 2g of maltose, 1g of dipotassium phosphate, 1g of zinc sulfate heptahydrate, 0.5g of sodium chloride, 0.5g of magnesium sulfate heptahydrate, 0.1g of ferrous sulfate heptahydrate, 0.1g of potassium dihydrogen phosphate, 15-20g of agar powder, 1L of distilled water, pH of 6.0-6.5, 0.1Mpa and sterilization at 121 ℃ for 15 min). In order to avoid Maillard reaction between the soybean protein and maltose after high temperature sterilization, the two are separately sterilized and then mixed uniformly. Culturing at 30 deg.C in dark, selecting strains with high growth speed and good spore growth, inoculating to slant culture medium of soybean juice, and culturing at 30 deg.C in incubatorAfter the materials are cultured to be mature, the materials are placed in a refrigerator at 4 ℃ for storage. Subsequently, the selected strains are coated on a Chachi plate, cultured for 5h at 30-32 ℃, observed by a microscope, and screened according to the hypha form by combining with high-flux re-screening, so as to obtain a strain with long hyphae, which is named as Aspergillus oryzae (Aspergillus oryzae) ZA216, and the hyphae length of the strain is shown in figure 1. A schematic representation of Aspergillus oryzae (Aspergillus oryzae) ZA216 is shown in FIG. 2.
The deposited information on Aspergillus oryzae (Aspergillus oryzae) ZA216 is:
aspergillus oryzae (Aspergillus oryzae) ZA216 was deposited at 18.5.2021 at the Collection of microorganisms and strains of Guangdong province under the accession number GDMCC NO:61673, deposit address: guangzhou city, first furious Zhonglu No. 100 large building No. 59.
Example 2 cultivation of Aspergillus oryzae (Aspergillus oryzae) ZA216
Inoculating Aspergillus oryzae (Aspergillus oryzae) ZA216 to bean juice slant for activation, culturing at 30 deg.C in dark for 4 days, and storing in 4 deg.C refrigerator after maturation. Inoculating mature slant onto triangular flask culture medium respectively for propagation (wherein the triangular flask culture medium is prepared from testa Tritici, semen glycines powder, flour and water; conventional method is adopted for process control), and detecting the resultant strain index (by conventional detection method), with the detection results shown in Table 1.
TABLE 1 Aspergillus oryzae (Aspergillus oryzae) ZA216 Erlenmeyer flask quality test for finished koji
Remarking: the indexes of the control strain Shanghai brewing 3.042 are 1.00, and the value of the mutagenized Aspergillus oryzae ZA216 is converted into the corresponding ratio.
EXAMPLE 3 investigation of the Properties of Aspergillus oryzae (ZA 216) Filter oil
Inoculating Aspergillus oryzae (Aspergillus oryzae) ZA216 to bean juice slant for activation, culturing at 30 deg.C in dark for 4 days, and storing in 4 deg.C refrigerator after maturation. Inoculating a mature inclined plane to soybean and wheat according to a mass ratio of 7:3, preparing the finished koji of Aspergillus oryzae (Aspergillus oryzae) ZA216 by a conventional method without special treatment in the conventional koji-making process, namely completely and normally operating on the basis of the Hu brewing 3.042 koji-making process.
100g of the yeast material prepared above is taken, added with 1.9 times of saline water, stirred evenly, kept stand for 2h and filtered. The results of comparing the differences between the soy mash sensory and the filter oil prepared from Aspergillus oryzae (Aspergillus oryzae) ZA216 and the strain Hu brewing 3.042 are shown in Table 2.
TABLE 2 oil filtration measurement of Aspergillus oryzae (Aspergillus oryzae) ZA216
| Sample/index | Oil filtering quantity |
| Shanghai brewing 3.042 | 1.00 |
| Aspergillus oryzae (Aspergillus oryzae) ZA216 | 0.89 |
Remarking: the indexes of the control strain Shanghai brewing 3.042 are 1.00, and the value of the mutagenized Aspergillus oryzae ZA216 is converted into the corresponding ratio.
From the viewpoint of oil filterability, 2h of the Aspergillus oryzae 3.042 strain in Shanghai is faster in oil filtration, and the Aspergillus oryzae (Aspergillus oryzae) ZA216 is slower in oil filtration, which shows that the Aspergillus oryzae (Aspergillus oryzae) ZA216 koji has better water absorbability. Aspergillus oryzae (Aspergillus oryzae) ZA216 has long hypha, good water retention, heavy yeast material, and easy sedimentation and water absorption. The Shanghai brewing 3.042 strain has short hypha, poor wrapping property of yeast, much water loss, light weight, easy floating on the surface and little water absorption. Because Aspergillus oryzae ZA216 has good water absorption, enzyme in the koji can be diffused and act on the particles of the soy sauce mash, thereby decomposing the nutrient components in the raw materials better and improving the utilization rate of the raw materials.
Example 4 use of Aspergillus oryzae (ZA 216) in Soy sauce production
Inoculating a mold starter (obtained by culturing Aspergillus oryzae 3.042 strain by conventional culture method) prepared from Aspergillus oryzae ZA216 into a culture medium prepared from soybean and wheat according to the mass ratio of 7:3 preparing a koji material, adopting a disc koji making machine to make koji according to a conventional method, completely and normally producing the koji without special treatment in the conventional koji making production process, namely on the basis of the koji making production process of Huniang 3.042, obtaining the finished koji of Aspergillus oryzae (Aspergillus oryzae) ZA216, and detecting the indexes as shown in table 3.
Adding 2 times of saline water into the disc koji making to perform high-salt dilute natural fermentation, wherein the fermentation period is within the range of 60-90 days according to the external environment, and performing 2 batches of high-salt dilute natural fermentation. After the fermentation is finished and the squeezing is carried out, indexes such as amino acid nitrogen, total nitrogen and the like are detected, and the detection results are shown in table 4. The results show that the indexes of amino acid nitrogen and total nitrogen are improved. The aroma and the mouthfeel on the sense are both obviously superior to those of 3.042 strain soy sauce, wherein the aroma is thick and rich, the delicate flavor is prominent, and the mouthfeel is mellow and long (as shown in figure 3).
TABLE 3 Aspergillus oryzae (Aspergillus oryzae) ZA216 fermentation starter propagation index detection
Remarking: the indexes of the control strain Shanghai brewing 3.042 are 1.00, and the value of the mutagenized Aspergillus oryzae ZA216 is converted into the corresponding ratio.
TABLE 4 Aspergillus oryzae (Aspergillus oryzae) ZA216 fermented Soy sauce index assay
Remarking: the indexes of the control strain Shanghai brewing 3.042 are 1.00, and the value of the mutagenized Aspergillus oryzae ZA216 is converted into the corresponding ratio.
Example 5 genetic stability test of Aspergillus oryzae (Aspergillus oryzae) ZA216
The selected Aspergillus oryzae (Aspergillus oryzae) ZA216 strain was continuously passaged on a soybean milk slant medium for 10 generations, the growth of the strain was observed and recorded for each generation, and the 1 st, 5 th and 10 th generation slant seeds were cultured and fermented as described in example 3. The amino acid nitrogen and total nitrogen contents of the fermented soy sauce were measured to determine the genetic stability of Aspergillus oryzae ZA216, and the measurement results are shown in Table 5.
TABLE 5 Aspergillus oryzae (Aspergillus oryzae) ZA216 genetic stability assay results
| Passage number | Amino acid nitrogen | Total nitrogen |
| Shanghai brewing 3.042 | 1.00 | 1.00 |
| ZA216 generation 1 | 1.06 | 1.02 |
| |
1.06 | 1.02 |
| ZA216 generation 10 | 1.06 | 1.02 |
Remarking: the indexes of the control strain Shanghai brewing 3.042 are 1.00, and the value of the mutagenized Aspergillus oryzae ZA216 is converted into the corresponding ratio.
As can be seen from Table 5, aspergillus oryzae ZA216 screened by the present invention has better genetic stability, and is favorable for the subsequent fermentation of soy sauce.
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, and 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 (9)
1. Aspergillus oryzae (Aspergillus oryzae) ZA216, deposited at the Guangdong province Collection of microorganisms and strains at 2021, 5, 18, with the deposit number GDMCC NO:61673, the preservation address is No. 59 of the Zhonglu-100 Jie-da, guangzhou city.
2. Use of Aspergillus oryzae (ZA 216) according to claim 1 in the fermentation of spices.
3. Use according to claim 2, wherein the seasoning is soy sauce or sauce.
4. Use of Aspergillus oryzae (ZA 216) according to claim 1 for the preparation of koji.
5. Use according to claim 4, wherein the koji obtained from koji making is used to prepare soy sauce or sauce.
6. A koji, characterized in that the koji is prepared by inoculating Aspergillus oryzae (ZA 216) according to claim 1 into a koji material prepared from soybean/defatted soybean and wheat.
7. Use of Aspergillus oryzae (ZA 216) according to claim 1 for improving the utilisation of raw materials in soy sauce or sauce fermentation.
8. A method for producing a soy sauce or a soy sauce, characterized in that Aspergillus oryzae (ZA 216) according to claim 1 is used as a fermentation starter.
9. A soy sauce or a paste obtained by using Aspergillus oryzae (ZA 216) as claimed in claim 1 as a fermentation starter.
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