72 results about "Exogenous enzymes" patented technology

The invention discloses a yeast beta-glucan preparation method based on high pressure-microjet homogenization treatment, and belongs to the technical field of zymosan extraction. The method comprises the following steps: carrying out high pressure-microjet homogenization pretreatment on active dry yeast, then carrying out autolysis treatment, then extracting beta-glucan by an acid-alkali method, and finally carrying out freeze drying to obtain a high-purity product. The core of the method is that the yeast cell walls are subjected to physical wall breaking by adopting a high pressure-microjet homogenization technology, and the extraction rate and the purity of the beta-glucan are remarkably improved by combining with an optimized acid-alkali treatment process. Compared with a traditional enzymolysis method, the method has the advantages that damage of exogenous enzyme to the beta-glucan structure is avoided, the purity of the obtained product reaches 84% or above, the content of protein impurities is lower than 0.2%, a new way is provided for efficient utilization of yeast cell wall resources, and the method is suitable for the fields of food, medicine and health care products.

The invention relates to a preparation method of a double-bacterium fermented opuntia ficus-indica fermentation extract and application of the double-bacterium fermented opuntia ficus-indica fermentation extract in cosmetics, and belongs to the technical field of cosmetics. The invention provides a preparation method of a double-bacterium fermented opuntia ficus-indica fermentation extract and application of the double-bacterium fermented opuntia ficus-indica fermentation extract to cosmetics, and the preparation method roughly comprises the following steps: preparing opuntia ficus-indica stem slurry and rice milk, mixing the slurry and the rice milk, performing double-bacterium fermentation, centrifuging, retaining residues, and finally enriching supernate. According to the invention, food-grade opuntia ficus stem and rice are used for fermentation, and aspergillus oryzae and saccharomycetes are used for combined fermentation, so that the introduction of exogenous enzyme is avoided, macromolecular substances are fully decomposed through synergistic metabolism, the release rate of micromolecular active ingredients is increased, and finally, the effects of moisturizing, oxidation resistance, wrinkle resistance and the like of the product are further improved through enrichment.

The invention relates to a saccharomycetes strain with high yield of esterifying enzyme and application thereof, the classification name of the saccharomycetes strain is Wickerhamomyces anomalus, the saccharomycetes strain is preserved in China General Microbiological Culture Collection Center on March 21, 2025, the preservation number is CGMCC 33925, and the address of the preservation unit is No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The strain screened by the invention has the ability of high yield of esterifying enzyme, and can synchronously catalyze synthesis of a plurality of key esters such as ethyl acetate, ethyl hexanoate, ethyl lactate and the like. Through the synergistic effect of polyesters, the flavor level of the liquor body is richer, and the liquor body is close to the composite aroma characteristic of the traditional solid fen-flavor liquor. The strain is naturally adapted to brewing of fen-flavor liquor and intensified application of medium and low temperature yeast, exogenous enzyme dependence is reduced, and the esterification capacity of yeast is improved.

The application discloses a method for preparing lactic acid by using a combined strain, and belongs to the technical field of microbial fermentation. Bacillus subtilis The combined strain comprises Bacillus subtilis Lactiplantibacillus pentosus LB9.The method for preparing lactic acid comprises the following steps: (1) preparing raw materials and strains, wherein the raw materials are white spirit lees; (2) mixing Bacillus subtilis and Lactobacillus pentosus after being activated respectively to form a combined strain, inoculating the combined strain into a culture medium taking the white spirit lees as the only fermentation substrate and without adding exogenous enzyme preparations and exogenous auxiliary materials, and performing fermentation; in the fermentation process, a staged oxygen supply strategy is adopted, namely, performing aerobic fermentation first and then performing anaerobic fermentation; and (3) collecting lactic acid from a fermentation product.The application realizes high-value utilization of lignocellulose waste, effectively coordinates metabolic conflicts among strains, and solves the problem of efficient conversion of lignocellulose lees into lactic acid.

A multi-part processed human amniotic composition configured to treat Peyronie's disease in a subject in need thereof by intracorporeal injection of the composition into the corpus cavernosum of the subject to reduce plaque size associated with Peyronie's disease. The multi-part processed human amniotic composition includes a micronized human amnion composition; and an aqueous human amniotic fluid filtrate configured to reconstitute and suspend the micronized human amnion composition therein. In certain aspects, the multi-part processed human amniotic compositions are not processed with exogenous enzymes during production thereof and do not include exogenous enzymes, such as collagenase, added thereto.

The application discloses a high-activity UGT1A1 enzyme based on tree shrew and a feed composition for enhancing alcohol metabolism. The catalytic efficiency of the recombinant tree shrew UGT1A1 enzyme on ethanol is significantly higher than that of human isozyme. The core of the application is to provide a complex feed system containing the enzyme, which solves the key bottleneck of poor stability and low delivery efficiency of exogenous enzymes in application through the synergistic effect of enzyme stabilizers, cell penetration enhancers, enteric coatings and coenzyme supply agents. Animal experiments show that the composition can safely and effectively accelerate in vivo alcohol elimination and reduce blood ethanol concentration, and can be used for developing alcoholism products, acute alcoholism treatment drugs and alcoholic liver disease prevention preparations.

The utility model relates to the field of oolong tea rocking barrels, and discloses an oolong tea adding external source enzymolysis rocking barrel which comprises a barrel body, a gear ring is arranged on the outer wall of the right end of the barrel body, the left side and the right side of the barrel body are both rotationally connected with fixing discs, a motor is installed on the right side of the fixing disc on the right side, and the driving end of the motor is fixedly connected with a rotating rod. A stirring rod is arranged in the barrel body, connecting buckles are arranged on the left side of a rotating rod and the right side of the stirring rod, a first belt wheel is arranged on the outer wall of the rotating rod, a fixing frame is fixedly connected to the front side of a fixing disc on the right side, a connecting rod is rotatably connected to the interior of the fixing frame, and a gear is fixedly connected to the left side of the connecting rod; the right side of the connecting rod is fixedly connected with a second belt wheel. According to the oolong tea enzymolysis device disclosed by the utility model, the motor, the belt pulley I, the gear, the gear ring and the stirring rod are matched with one another, so that the box body and the stirring rod rotate in opposite directions, and oolong tea is subjected to more comprehensive enzymolysis, so that the working efficiency is improved.

This invention discloses a method for improving the flavor of sweet rice wine through low-temperature fermentation and its brewing technique, relating to the field of winemaking technology. First, a short-term temperature increase (28-30℃) is implemented in the early stages of fermentation to create an optimal environment for efficient saccharification and flavor precursor accumulation in Rhizopus. Subsequently, a low-temperature stage (15-20℃) is initiated, under which slow primary fermentation and flavor maturation, primarily driven by yeast, take place. The "staged temperature-increase saccharification + low-temperature fermentation" process provided by this invention does not rely on psychrophilic strains or exogenous enzyme additives; it can be achieved through optimization of process parameters. This method not only successfully solves the core problem of insufficient saccharification kinetics at low temperatures, ensuring the smooth progress of the fermentation process, but also significantly improves the sensory quality of the final sweet rice wine (more complex and harmonious flavor), as well as the content of umami and sweet amino acids. Furthermore, this process is highly controllable and suitable for large-scale production, providing a novel technological path for the green and efficient brewing of high-quality sweet rice wine.

The invention discloses an enzyme-activated fluorescent molecular precursor as well as a preparation method and application thereof. The enzyme-activated fluorescent molecular precursor has a chemical structure as shown in a formula I which is described in the specification. The invention develops an enzyme-activated fluorescent molecular precursor which can be used for real-time imaging of tumors by targeting exogenous enzyme to the tumors and specifically activating a probe at the tumors; according to the invention, the exogenous enzyme is introduced as an activation trigger, and normal cells do not have the exogenous enzyme, so that the problem of co-expression of the endogenous enzyme in non-tumor tissues can be avoided, background fluorescence signals can be reduced to the greatest extent, and the imaging specificity and accuracy are remarkably improved.

The present disclosure concerns a process for fermenting a biomass with a reduced dose of a purified exogenous enzyme (which can be, for example a purified exogenous glucoamylase). The process comprises contacting a biomass (which may comprise starch) with a recombinant yeast host cell. The recombinant yeast host cell has a genetic modification for expressing a heterologous polypeptide having starch or dextrin hydrolase activity (which may be, for example, from a glucoamylase). The nucleic acid molecule encoding the heterologous polypeptide having starch or dextrin hydrolase activity comprises allowing the secretion of the heterologous polypeptide.

The present invention is directed to a yeast strain, or strains, secreting a full suite, or any subset of that full suite, of enzymes to hydrolyze corn starch, corn fiber, lignocellulose, (including enzymes that hydrolyze linkages in cellulose, hemicellulose, and between lignin and carbohydrates) and to utilize pentose sugars (xylose and arabinose). The invention is also directed to the set of proteins that are well expressed in yeast for each category of enzymatic activity. The resulting strain, or strains can be used to hydrolyze starch and cellulose simultaneously. The resulting strain, or strains can be also metabolically engineered to produce less glycerol and uptake acetate. The resulting strain, or strains can also be used to produce ethanol from granular starch without liquefaction. The resulting strain, or strains, can be further used to reduce the amount of external enzyme needed to hydrolyze a biomass feedstock during an Simultaneous Saccharification and Fermentation (SSF) process, or to increase the yield of ethanol during SSF at current saccharolytic enzyme loadings. In addition, multiple enzymes of the present invention can be co-expressed in cells of the invention to provide synergistic digestive action on biomass feedstock. In some aspects, host cells expressing different heterologous saccharolytic enzymes can also be co-cultured together and used to produce ethanol from biomass feedstock.

The invention provides non-sugar-tolerant beta-glucosidase, an expression vector and application of the non-sugar-tolerant beta-glucosidase in improvement of cellulose degradation capacity, and belongs to the technical field of genetic engineering. According to the invention, the non-sugar tolerance type beta-glucosidase BGLC27 is obtained from streptomyces decomposer C27, and the amino acid sequence of the BGLC27 is as shown in SEQ ID No. 1. According to the invention, the BGLC27 is used as an exogenous enzyme and is added into a degradation system of cellulose degradation bacteria, and research finds that after the BLGC27 is exogenously added into the compound bacteria system, the cellulose degradation capability can be obviously improved.

The present application relates to the field of food technology, and discloses a low-bitterness wolfberry original syrup and a preparation process thereof. The original syrup is prepared from the following raw materials in parts by weight: 80-90 parts of fresh wolfberry fruit, 5-10 parts of germinated fava, 5-10 parts of germinated barley, 0.3-0.6 parts of medium-chain triglyceride, 1.0-2.0 parts of concentrated red date juice, 0.05-0.1 parts of ascorbic acid, 0.02-0.05 parts of citric acid, and 0.01-0.05 parts of saltwort extract. The present application efficiently reduces the bitterness of the wolfberry original syrup through a full natural biological transformation pathway without adding exogenous enzyme preparations or introducing chemical adsorbents, fundamentally avoiding chemical addition and clean label problems, and simultaneously achieving the synchronous transformation of various types of bitter substances in wolfberry. The preparation process of the present application completely retains the heat-sensitive functional factors and fresh fruit aroma of wolfberry, and the nutritional integrity and flavor reduction degree of the product are significantly better than those of similar products on the market.

The invention discloses a multifunctional active peptide from mackerel viscera and application of the multifunctional active peptide. The mackerel viscera extract is prepared by adopting a single exogenous enzyme hydrolysis method, the extract has small molecule peptides with ACE inhibitory activity, three ACE inhibitory peptides are obtained by further identification and screening, the amino acid sequences are respectively GPPVPGP, GPMGPMGP and SPGPVGMP, and the half inhibitory concentration IC50 values of the ACE inhibitory peptides are respectively 80.84 + / -1.12 mu g / mL, 177.41 + / -3.26 mu g / mL and 163.25 + / -3.46 mu g / mL. Wherein the heptapeptide GPPVPGP with the optimal activity also has the function of removing DPPH free radicals and hydroxyl free radicals, and can be used for adjuvant therapy for reducing hypertension or applied to other fields.

The invention discloses a black tea processing method for directionally releasing sweet flower fragrance characteristic components based on exogenous enzyme targeted regulation and control of glucoside hydrolysis. The black tea processing method comprises the following steps: withering fresh leaves, performing enzymatic treatment, performing gradient rolling, performing temperature-controlled humidity-controlled fermentation, and performing gross fire-spreading airing-complete fire staged drying on the fermented tea leaves, an enzyme solution sprayed for enzymatic treatment comprises one or more of an alpha-galactosidase solution with the activity being 0.5 to 0.6 U / mL, a beta-glucosidase solution with the activity being 0.6 to 0.7 U / mL, a cellulase solution with the activity being 200 to 210 U / mL and a papain solution with the activity being 1600 to 1700 U / mL; the sweet flower fragrance characteristic components comprise at least one of beta-myrcene, junifolene, citral, linalool, methyl salicylate, geraniol and nerol. The method solves the industrial problems of insufficient release of sweet flower fragrance characteristics, poor process stability and high dependence degree of raw materials in traditional black tea processing.

The invention provides a biosynthesis method for efficiently producing aromatic non-natural amino acid and site-specific modified recombinant protein, an engineering strain and application of the engineering strain. Specifically, the invention provides an engineering bacterium, and genes of one or more exogenous enzymes selected from the following groups are introduced into the engineering bacterium: threonine aldolase, threonine deaminase and transaminase; and threonine aldolase, threonine deaminase and transaminase are simultaneously expressed in the engineering bacterium. The engineering bacterium disclosed by the invention can be used for efficiently synthesizing aromatic non-natural amino acid through whole-cell catalysis or fermentation conversion. More importantly, the synthetic route provided by the invention can be synergistically coupled with a gene codon extension technology in the same engineering bacterium, and is used for high-efficiency expression of recombinant target protein modified by aromatic non-natural amino acid at a fixed point.

The invention discloses a method for increasing the content of sulforaphane in broccoli through magnetic field assisted blanching, and belongs to the technical field of food processing. The method for improving the content of sulforaphane in broccoli through magnetic field assisted blanching comprises the following steps: (1) preparing raw materials; (2) magnetic field assisted blanching pretreatment; (3) cooling and structure keeping treatment; (4) carrying out enzymatic incubation based on a magnetic field-assisted blanching regulation state; and (5) obtaining the broccoli product with high sulforaphane content. Exogenous enzymes, chemical reagents or additives are not introduced in the process, and the activity of endogenous myrosinase and the catalytic hydrolysis path of the endogenous myrosinase are regulated by synchronously applying a physical regulation means in the blanching pretreatment process, so that the generation level of sulforaphane in a treated sample is improved. The method is suitable for a food processing system, has the advantages of mild process conditions, strong controllability, high safety and easiness in industrial amplification, and has a wide application prospect in the field of food processing.

The invention belongs to the field of agriculture, and particularly discloses a biostimulant compound fertilizer containing sea cucumber peptide and a preparation method thereof, the biostimulant compound fertilizer containing sea cucumber peptide comprises the following components by weight: 5-15 parts of a sea cucumber peptide compound metabolite, 10-20 parts of humic acid, 10-15 parts of potassium fulvate, 40-60 parts of a nitrogen-phosphorus-potassium compound fertilizer, 1-5 parts of trace elements, 5-10 parts of alginic acid, and 5-10 parts of chitosan, the sea cucumber peptide composite metabolite is prepared through microbial fermentation and comprises antibacterial peptide, polyglutamic acid and oligosaccharide. The sea cucumber peptide composite metabolite rich in antibacterial peptide-polyglutamic acid-oligosaccharide is efficiently prepared through a three-stage synergistic enzymolysis process and a microbial symbiotic fermentation technology, a functional synergistic system is formed by combining components such as humic acid and chitosan, the enzymolysis efficiency is remarkably improved, an exogenous enzyme is replaced with a viscera natural enzyme source, the production cost is reduced, and the method is suitable for industrial production. And the enzymolysis efficiency is improved by combining a body wall pretreatment technology.

This invention discloses a fully solid-state fermentation process for Astragalus membranaceus vinegar, specifically relating to the field of solid-state vinegar brewing technology. The process uses sorghum, rice husks, rice bran, wheat bran, Astragalus membranaceus, and Daqu (a type of starter culture) as raw materials, without adding any exogenous enzymes or chemical products. The process includes the following steps: S1. Astragalus membranaceus slices are diagonally sliced ​​and drained to a moisture content of 18%-22%; S2. Sorghum is steamed at normal pressure, with Astragalus membranaceus slices added after 20-25 minutes of steaming, and steaming continues for a total time of 40-60 minutes; S3. Sorghum undergoes variable-temperature alcoholic fermentation with rice bran, first-grain rice husks, and a portion of Daqu, without Astragalus membranaceus slices participating in this stage; S4. The mash is mixed with the remaining Daqu, wheat bran, second-grain rice husks, and softened Astragalus membranaceus slices under moist heat, and the moisture content is adjusted to 60%-65% before acetic acid fermentation, with the main fermentation temperature at 39℃-41℃; S5. The mash is aged in a sealed container with intermittent aeration; S6. Vinegar is obtained by gradient temperature leaching. This invention achieves simultaneous fermentation and self-degradation of Astragalus membranaceus fiber and in-situ conversion of its functional components through process timing control and physical parameter optimization.

The invention discloses a processing method for improving clarity of instant tea, and belongs to the technical field of deep processing of tea leaves, the method mainly comprises the following steps: extracting and roughly filtering tea leaves to obtain primary tea soup; adding a compound enzyme into the primary tea soup, carrying out enzymolysis treatment, and then carrying out enzyme deactivation; sequentially performing ceramic membrane microfiltration, ultrafiltration membrane fine filtration and nanofiltration membrane concentration on the treated tea soup; quickly cooling the obtained concentrated solution, standing and aging at low temperature, and centrifuging at high speed to obtain supernatant liquid; and finally, drying to obtain the instant tea powder. According to the method, various substances causing turbidity in tea soup are systematically removed through the synergistic effect of exogenous enzymolysis, three-stage membrane integrated purification and low-temperature physical coagulation acceleration, and the method has the beneficial effects that the final clarity and cold water solubility of an instant tea product can be effectively improved, and the storage stability of the product is improved.

The application provides a vinegar residue-based single-cell protein feed and a preparation method and application thereof, and belongs to the technical field of feed preparation.The application uses vinegar residue as raw material, adopts solid-state fermentation of Aspergillus, Candida utilis and Saccharomyces cerevisiae in combination with composite enzymes, utilizes the synergistic effect among the strains and the enzymolysis effect of the exogenous enzymes, efficiently degrades the fibrous substances in the vinegar residue, and converts the fibrous substances into microbial proteins, so that the crude protein content of the prepared vinegar residue-based single-cell protein feed is increased from 5% to 8% of the raw material to 20% to 25%, and the total fibrous substance degradation rate is greater than or equal to 60%. The vinegar residue-based single-cell protein feed produced by the application has good feeding value, can effectively replace part of the protein raw materials such as soybean meal, improves the production performance of beef cattle, reduces the breeding cost, realizes high-value utilization of vinegar residue resources, and has good popularization and application prospect.

The present invention relates to the field of genetic engineering technology, and discloses a plasmid-free genetically engineered bacterium with high yield of L-2-amino butyric acid and its application. The plasmid-free genetically engineered bacterium for producing L-2-amino butyric acid provided by the present invention is obtained by modifying Escherichia coli THR as a chassis cell through metabolic engineering and gene editing technology. This genetically engineered bacterium strain can use glucose as a raw material and obtain L-2-amino butyric acid through fermentation culture. Without adding exogenous enzyme activity by plasmids, the batch-fed fermentation yield in a 5L fermenter reaches 13.81 g / L. It has low fermentation cost, high product concentration, no by-products affecting, and the product is easy to purify, and is suitable for industrial applications.

Provided in the present invention are a biosynthesis method for efficiently producing aromatic unnatural amino acids and site-specifically modified recombinant proteins, an engineered strain, and use thereof. Specifically, the present invention provides an engineered bacterium. Genes of exogenous enzymes, threonine aldolase and threonine deaminase, are introduced into the engineered bacterium, and the engineered bacterium expresses both the threonine aldolase and the threonine deaminase. The engineered bacterium of the present invention enables efficient synthesis of aromatic unnatural amino acids by means of whole-cell catalysis or fermentation transformation. The synthetic route of the present invention can be synergistically coupled with gene codon extension technology in the same engineered bacterium for efficient expression of recombinant target proteins site-specifically modified with aromatic unnatural amino acids.

A subject-matter of the present invention is a method for the dyeing of keratin fibres employing a composition C comprising: (i) indican of formula (I) and / or a powder containing it; (ii) at least one exogenous enzyme of hydrolase type; (iii) at least one exogenous enzyme of oxidase type; and (iv) water.

The invention discloses coix seed whisky and a preparation method thereof, and relates to the technical field of fermentation engineering. Comprising the following steps: treating raw materials; mash preparation and saccharification; a fermentation process; a primary distillation process; a secondary distillation process; a barrel aging process; and filling. The core of the method is that an innovative process of partially frying ungerminated coix seed raw materials in combination with synergistic saccharification of exogenous enzyme and malt is adopted, and a brand new process route of'relative low-temperature gelatinization-enzymatic saccharification-non-filtration direct fermentation distillation 'is constructed. Meanwhile, a partial coix seed stir-frying process, malt auxiliary saccharification and Mongolian oak barrel aging combined herbal pretreatment process are further innovatively introduced, and technical support is provided for whisky localization innovation.

The application discloses a construction method and application of Corynebacterium glutamicum for synthesizing shinorine, and belongs to the field of genetic engineering and fermentation technology. The Corynebacterium glutamicum ATCC 13032 is used as a starting strain, a competitive pathway gene is knocked out, an optimal exogenous enzyme is screened, and a key enzyme gene cluster is integrated in multiple copies on a genome, so that the synthesis yield of shinorine in the microbial cell is improved, and the strain can accumulate 0.63 g / L of shinorine in 72 hours in a flask. In fed-batch fermentation in a 50 L fermenter, the final yield of shinorine can reach 4.3 g / L.

This invention discloses a method for the targeted conversion and synthesis of rare ginsenosides Rg3 and Rh2. Specifically, fresh Panax notoginseng leaves are microwave-dried for 1-3 minutes to inactivate their endogenous β-glucosidase. The dried Panax notoginseng leaves are extracted with anhydrous methanol, and the extract is resuspended in an acetate-sodium acetate buffer solution after drying. α-arabinofuranase, β-xylosidase, and β-glucosidase are added to the resuspended solution, and an enzymatic reaction is carried out at 35-40℃ to directionally convert the original ginsenosides into rare ginsenosides Rg3 and Rh2. This method can achieve the inactivation of endogenous enzymes and the stable retention of original saponins in Panax notoginseng leaves in a very short time. Compared with the natural drying combined with exogenous enzymes, the conversion ratios of Rg3 and Rh2 are increased by 3.14 and 2.4 times, respectively. Compared with fresh Panax notoginseng leaves, the contents of Rg3 and Rh2 are increased by 20.6 and 8.7 times, respectively. This method is suitable for the high-value utilization of Panax notoginseng leaf waste resources and the large-scale preparation of rare saponins.

This application relates to a hygroscopic yeast extract and its preparation method. The preparation method comprises the following steps: preparing yeast cell walls into a yeast cell wall suspension; adding active yeast to the yeast cell wall suspension to autolyze the yeast cell wall with endogenous enzymes to produce an autolysis product; adding papain, aminopeptidase, and flavor neutral protease to the autolysis product to hydrolyze the product with exogenous enzymes to produce an enzymatic hydrolysis product; and separating the hydrolysis supernatant from the enzymatic hydrolysis product to produce a hygroscopic yeast extract. This method achieves deep hydrolysis of the yeast cell walls, and the resulting yeast extract not only has a good, rich flavor but also exhibits excellent hygroscopic resistance.

The invention belongs to the field of agricultural microorganism and waste resource utilization, and discloses a functional complementary flora and cellulase synergistic straw decomposition agent and application thereof. The decomposition agent is prepared by scientifically compounding a complex microbial community consisting of bacillus proteolyticus WYX-67, P.avenae WYX-3 and paenibacillus polysaccharificus WYX-166 with industrial cellulase, and the complex microbial community consists of bacillus proteolyticus WYX-67, P.avenae WYX-3 and paenibacillus polysaccharificus WYX-166. The technical bottlenecks of product inhibition, enzyme activity attenuation, slow start and the like in application of a single preparation are effectively overcome through the'quick start 'effect of exogenous enzyme and'continuous relay' degradation of microbial flora. Test data show that the decomposition rate and the organic carbon mineralization ability of the wheat straw can be remarkably improved, and a brand new technical scheme is provided for efficient and environment-friendly treatment of the straw returned to the farmland.

The invention provides a soy sauce brewing method and soy sauce, the soy sauce brewing method adopts an ancient-process koji making and an ancient-process jar fermentation brewing, innovatively combines a gradient ratio hyperbolic process, double fermentation and double sun-curing, does not need to additionally add materials such as an exogenous enzyme preparation and caramel color, belongs to single-strain koji making, and is simple in process, low in cost and easy to popularize. The koji making technology is relatively simple, long-period brewing is conducted in the natural environment, the Maillard reaction is more sufficient, and the produced ancient-method soy sauce is thicker in body, more fragrant and sweet in taste and richer in sauce fragrance.