CN102115734B - Special compound enzyme for yeast hydrolysis and preparation method thereof - Google Patents
Special compound enzyme for yeast hydrolysis and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a special compound enzyme for yeast hydrolysis and a preparation method thereof. The compound enzyme comprises the following enzyme components in parts by weight: 1-30 parts of beta-glucanase, 1-30 parts of mannase, 1-30 parts of saccharifying enzyme, 15-65 parts of endoenzyme, 1-10 parts of exoenzyme, 1-10 parts of flavourzyme and 1-10 parts of phosphodiesterase. The preparation method comprises the following steps: respectively fermenting, extracting, freeze-drying and pulverizing the beta-glucanase, mannase, saccharifying enzyme, endoenzyme, exoenzyme, flavourzyme and phosphodiesterase, and evenly mixing according to the proportion designed in the formula to obtain the special compound enzyme for yeast hydrolysis. Since multiple biological enzymes are coupled together for hydrolysis, the yeast hydrolysis time is shortened from 20-32 hours in the traditional way to 10-25 hours, the utilization ratio of the proteins is higher than 90%, the generation rate of amino acids is higher than 75%, the content of amino acids is higher than 0.83g/100ml, and the total nitrogen content on dry basis is higher than 12.0%.
Description
Technical field
The invention belongs to the zymin technical field, relate to a kind of yeast hydrolysis special composite enzyme and preparation method thereof.
Background technology
Yeast contains rich in protein, dextran, mannosans, VITAMIN, mineral substance and a large amount of gsh etc., also contain 4～15% degradable Yeast Nucleic Acid (RNA) in addition, pass through zymolysis technique, decompose yeast cells wall, stripping protein, proteolytic enzyme further resolves into protein little peptide, amino acid etc., phosphodiesterase decomposes Yeast Nucleic Acid again and generates flavour nucleotide materials such as (I+G), make yeast extract product, it not only has good trophism, but also has good health-care effect, it is a kind of seasoning that has, the condiment of three big functions such as nutrition and health care can substitute products such as existing monosodium glutamate, and market outlook are very huge.In natural condiment containing, yeast extract occupies critical role, generally replaces the meat hydrolyzate or replaces monosodium glutamate to make daily seasonings with yeast extract in country such as American-European-Japanese, and also alternative extractum carnis etc. is as the biological fermentation nutrition agent.
Beer of China and sugarcane yield are all at the forefront in the world, and byproduct beer yeast slurry after it is produced and cane molasses yeast provide abundant yeast raw material resources for zymic deep processing yeast extract product.China's cane molasses output reached 280～3,400,000 tons in 2009, and the output of beer yeast slurry is 50～600,000 tons, did the waste processing and caused the serious wasting of resources and environmental pollution, and the economic worth of waste yeast is not fully used for a long time.The huge yeast resource of exploitation and comprehensive utilization China has very significant social, economy and environmental benefit.
The production technique of yeast extract is autolysis method normally, three kinds of methods such as acid and alkali hydrolysis method and biological enzyme.Production time of autolysis method is long, it is easily smelly to be bacterial contamination, degree of hydrolysis is low, protein utilization rate is low; Problems such as the chemical acid alkali method exists saltiness height, weak flavor, nutritive loss is big, security is low; And biological enzyme has the reaction conditions gentleness, non-pollutant discharge, characteristics such as hydrolysis rate is fast, thorough enzymolysis, hydrolysis amino nitrogen content height, little, the bright local flavor of bitter taste are good.According to literature search, since nineteen ninety-three, units such as domestic northwest light industry institute, Wuxi Light Industry Univ., South China Science ﹠ Engineering University, Guangdong microbiological industry institute all make a search to yeast hydrolysis, yeast extract production etc., what adopt all is single or two kinds of biological enzyme hydrolysis that share such as papoid, neutral protease, Sumizyme MP, its degree of hydrolysis and amino nitrogen content are all lower, raw materials for production consume and the cost height, and delicate flavour and local flavor are general.A lot of researchs are also done to the yeast hydrolysis by countries such as external Japan and Denmark, and for example Denmark's promise believes that only company's using basic proteolytic enzyme or compound protease and food flavor enzyme are better in the yeast hydrolysis effect, but the price height.The yeast hydrolysis special composite enzyme of requirements such as in sum, existing market is badly in need of satisfying degree of hydrolysis height, amino nitrogen height very much comprehensively, bright local flavor is good, cost is low.
Summary of the invention
The technical problem that the objective of the invention is that the hydrolysis material that exists at the yeast hydrolysis is limited, sporoderm-broken rate is low, protein utilization is low, degree of hydrolysis is low, yield is low, the hydrolysis substance point of contact is not good etc. provides a kind of yeast hydrolysis special composite enzyme and preparation method thereof.
Technical scheme of the present invention is:
The inventor is on traditional autolysis method of further investigation and modern biological enzyme hydrolysis mechanism based, and the material molecular structure characteristics of combining yeast are studied the plurality of enzymes of energy hydrolysed leaven, have found new method and technology, have also opened up another kind of thinking.The yeast belong fungi, its cell comprises diversified biologically active substance, and protein content is up to 35～50%, and is all high than other animal proteinum raw materials such as beef, pork etc., contains a large amount of essential amino acids, and amino acid is formed rationally.Also contain abundant VITAMIN, mineral substance and a large amount of gsh, dextran and mannosanss etc. simultaneously, contain 4～15% Yeast Nucleic Acid (RNA) in addition, but these nutritive substances are present in the yeast cell mostly, and most protein are insoluble, can not ooze out from cell walls and cytolemma.So must destroy cell walls and cytolemma by scientific methods, make nutritive substance exposed with discharge, and make these materials change into effective constituent with delicious taste and nutrition.Take a broad view of existing yeast decomposition method, because the enzyme that they adopt is single, or restriction endonuclease, or excision enzyme or food flavor enzyme, the yeast material molecular structure there is not specific aim, multiple material composition plyability effectively with is not comprehensively acted on, thereby protelytic effect is relatively poor, major part can not become macromolecular Yeast protein the polypeptide and the amino acid of target fully, and nutritive ingredient is incomplete, does not more have good mouthfeel and local flavor.And the present invention is under the condition of the temperature that sets, pH value and time, opened the complex construction of yeast cells wall by couplings such as the beta-glucanase in the yeast hydrolysis special composite enzyme, mannase, saccharifying enzyme, cut off wherein stable physical chemistry key, make the protein molecular structure become loose, nonpolar albumen group comes out, and helps the protein incision enzyme effect and accelerates the Yeast protein hydrolysis; In addition, under the restriction endonuclease effect in yeast hydrolysis special composite enzyme, cut off the peptide bond of Yeast protein macromole peptide chain inside, generate intermediate products such as less peptone of molecular weight and polypeptide, by the excision enzyme in the yeast hydrolysis special composite enzyme free C-terminal and N-terminal in the polypeptide are cut off peptide bond hydrolysis one by one then and generate materials such as low little peptide of molecule and biological activity amino acid, be aided with the auxiliary modification of food flavor enzyme, nutritious to obtain, degree of hydrolysis is high, the yeast hydrolysis prods of functional excellent and raciness.The main fragrant delicate flavour that generates 5 ' one Nucleotide taste compounds regulation and control yeast hydrolyzed solution reduces bitter taste and makes the taste of Yeast protein hydrolysis prods better under the effect of the phosphodiesterase of Yeast Nucleic Acid (RNA) in yeast hydrolysis special composite enzyme, and fragrance is stronger.
Below be the chemical equation of the enzymolysis mechanism of concrete reaction:
The present invention is achieved in that
A kind of yeast hydrolysis special composite enzyme, comprise beta-glucanase, mannase, saccharifying enzyme, restriction endonuclease, excision enzyme, food flavor enzyme and phosphodiesterase, evenly obtain yeast hydrolysis special composite enzyme by the formulating of recipe mixed, its composition and parts by weight thereof are: beta-glucanase is 1～30 part, and mannase is 1～30 part, and saccharifying enzyme is 1～30 part, restriction endonuclease is 15～65 parts, excision enzyme is 1～10 part, and food flavor enzyme is 1～10 part, and phosphodiesterase is 1～10 part.
The preparation method of yeast hydrolysis special composite enzyme, be earlier with beta-glucanase, mannase, saccharifying enzyme, restriction endonuclease, excision enzyme, food flavor enzyme and phosphodiesterase respectively by fermentation, extraction, freeze-drying, pulverizing, evenly obtain yeast hydrolysis special composite enzyme by the formulating of recipe mixed then, its operational process of craft is:
(1) preparation of beta-glucanase: with soybean or dregs of beans or wheat bran is raw material, adopts viride (Trichoderma viride) bacterial classification or aspergillus niger (Aspergillus niger) bacterial classification to prepare according to a conventional method, preserves standby;
(2) preparation of mannase: with soybean or dregs of beans or wheat bran is raw material, adopts viride (Trichoderma viride) bacterial classification or aspergillus niger (Aspergillus niger) bacterial classification to prepare according to a conventional method, preserves standby;
(3) preparation of saccharifying enzyme: with soybean or dregs of beans or wheat bran is raw material, adopts viride (Trichoderma viride) bacterial classification or aspergillus niger (Aspergillus niger) bacterial classification to prepare according to a conventional method, preserves standby;
(4) preparation of restriction endonuclease: be prepared as example with papoid, take off new fresh Chinese flowering quince syrup from pawpaw tree or fruit, earlier with tap water or distilled water or deionized water towards rare or dissolving pawpaw latex, extract by freeze thawing then, obtain papoid through micro-filtration, ultrafiltration and concentration, vacuum lyophilization, pulverizing again, preserve standby; The preparation of neutral protease, Sumizyme MP obtains through fermented extracted according to a conventional method;
(5) preparation of excision enzyme, with soybean or dregs of beans or wheat bran is raw material, pulverize, sterilization, add aspergillus oryzae (Aspergillus oryzae) or Rhizopus oryzae (Rhizopus oryzae), aspergillus niger (Aspergillus niger) bacterial classification, 30～50 ℃, 70～90% humidity condition bottom fermentation 1～5 day, extraction then, micro-filtration, ultrafiltration, freeze-drying, pulverizing obtained, and preserve standby;
(6) preparation of food flavor enzyme, with soybean or dregs of beans or wheat bran is raw material, pulverize, sterilization, add Mucor (Actinomucor elegans), aspergillus oryzae bacterial classifications such as (Aspergillus oryzae), 25～45 ℃, 70～90% humidity condition bottom fermentation 1～4 day, extraction then, micro-filtration, ultrafiltration, freeze-drying, pulverizing obtained, and preserve standby;
(7) preparation of phosphodiesterase, with soybean or dregs of beans or wheat bran is raw material, pulverize, sterilization, add Penicillium citrinum (Penicillium citrinum) bacterial classification, 25～45 ℃, 70～90% humidity condition bottom fermentation 1～4 day, extraction then, micro-filtration, ultrafiltration, freeze-drying, pulverizing obtained, and preserve standby;
(8), mix by optimization of C design composition and parts by weight thereof and to obtain yeast hydrolysis special composite enzyme with the above-mentioned beta-glucanase for preparing, mannase, saccharifying enzyme, restriction endonuclease, excision enzyme, food flavor enzyme and phosphodiesterase.
Above-described beta-glucanase, mannase, saccharifying enzyme, be to extract by viride (Trichoderma viride) bacterial classification or aspergillus niger (Aspergillus niger) strain fermentation to prepare, wherein the beta-glucanase enzyme activity is 1000～100000u/g, the mannase enzyme activity is 5000～200000u/g, and the glucoamylase enzyme vigor is 5000～200000u/g.
Above-described restriction endonuclease comprises the mixture of papain, neutral protease and Sumizyme MP, and blending ratio is 0～5: 0～5: 0～5, and its total enzyme activity is 10000～800000u/g.
Above-described excision enzyme, comprise that one or more strain fermentations in aspergillus oryzae (Aspergillus orvzae) or Rhizopus oryzae (Rhizopus oryzae), the aspergillus niger (Aspergillus niger) extract the biological enzyme for preparing, its enzyme activity is 1000～50000u/g.
Above-described food flavor enzyme comprises that one or both strain fermentations in Mucor (Actinomucor elegans), the aspergillus oryzae (Aspergillus oryzae) extract the biological enzyme for preparing, and its enzyme activity is 1000～50000u/g.
Above-described phosphodiesterase is to extract the phosphodiesterase that obtains from Penicillium citrinum (Penicillium citrinum) strain fermentation, and its enzyme activity is 1000～70000u/g.
Above-described yeast hydrolysis special composite enzyme, it is applied to the yeast hydrolysising condition: molasses yeast, cereuisiae fermentum, bread yeast or other processing waste yeasts are added water be transferred to 8～20% concentration, under the temperature of the pH value 4.5～7.0 and 45～70 ℃, press 0.1～0.6% of substrate weight and add product yeast hydrolysis special composite enzyme of the present invention, enzymolysis 10～25 hours, be warmed up to then more than 85 ℃ and continue 15 minutes～1 hour the hydrolyzed solution enzyme that goes out, separate again, concentrate, dry, obtain yeast extract product at last.
Above-described yeast hydrolysis special composite enzyme, be used for yeast deep processings such as molasses yeast, cereuisiae fermentum, bread yeast or other process for processing waste yeasts, press the hydrolysising condition hydrolysis, obtain the yeast hydrolyzed solution, detect the hydrolyzed solution aminoacids content greater than 0.83g/100ml, give money as a gift amino nitrogen greater than 6.73%, give money as a gift total nitrogen greater than 12.0%, and have very strong delicate flavour and strong sauce fragrance, good trophicity and food safety, do not need the outside Nucleotide materials such as (I+G) that adds, do not contain harmful material.
Above-described yeast hydrolysis special composite enzyme is applicable to the application of yeast deep processings such as molasses yeast, cereuisiae fermentum, bread yeast or other process for processing waste yeasts.
Above-described yeast hydrolysis special composite enzyme, its yeast hydrolysis prods can be used for aspects such as biological fermentation, seasonings, meat-based food processing, processing of aquatic products, condiment for instant noodles, puffed food and nutritional supplements.
Advantage of the present invention and positively effect:
1, the present invention is on the basis of traditional autolysis method and modern biological enzyme hydrolysis mechanism, the proteinic cellularstructure characteristics of combining yeast, extract beta-glucanase, mannase, the saccharifying enzyme that is fit to breaking yeast cellule membrane with viride (Trichoderma viride) bacterial classification or aspergillus niger (Aspergillus niger) strain fermentation, so that dissolve the yeast cell wall construction effectively, protein is effectively discharged.
2, the present invention selects the endo-protease mixture that is fit to the yeast hydrolysis from yeast structure of matter characteristics and enzymolysis mechanism angle sieve, more effectively the yeast protein of stripping behind broken wall is hydrolyzed into materials such as peptide class and amino acid.
3, the present invention selects for use aspergillus oryzae (Aspergilius oryzae) or Rhizopus oryzae (Rhizopus oryzae), aspergillus niger induction mutation of bacterium, fermentation, extraction, separation such as (Aspergillus niger) to obtain the proteolysis excision enzyme from the microorganism of the traditional soy sauce production of China, and excision enzyme decomposes materials such as small protein and polypeptide and becomes small peptide and amino acid to obtain high degree of hydrolysis and amino nitrogen.
4, the present invention selects for use Mucor (Actinomucor elegans), aspergillus oryzae induction mutation of bacterium, fermentation, extraction, separation such as (Aspergillus oryzae) to obtain the proteolysis food flavor enzyme from the microorganism of the traditional production of preserved beancurd of China, when making the yeast hydrolysis cut enzyme outside to decompose materials such as small protein and polypeptide and become small peptide and amino acid to obtain high degree of hydrolysis and amino nitrogen, be aided with the auxiliary modification of food flavor enzyme again, nutritious to obtain, degree of hydrolysis is high, the yeast hydrolysis prods of functional excellent and raciness.
5, the present invention is from yeast material molecular structure characteristics and enzymolysis mechanism angle, obtain phosphodiesterase from Penicillium citrinum (Penicillium citrinum) induction mutation of bacterium, fermentation, extraction, separation, can decompose Yeast Nucleic Acid (RNA) in the Yeast protein generates 5 '-Nucleotide (I+G) taste compound, make yeast hydrolysis prods delicate flavour better.
6, yeast hydrolysis special composite enzyme of the present invention is used for the yeast hydrolysis, technology is improved to a step enzymolysis process by two step of original cell wall breaking technology+protein hydrolysis process enzymolysis process, hydrolysis time reaches 32 hours by original twice in step sum total and foreshortens to 10～25 hours, improves hydrolysis efficiency.
7, yeast hydrolysis special composite enzyme of the present invention is used for the yeast hydrolysis, makes the yeast protein utilization ratio greater than 90%, and the amino acid production rate is more than 75%.
8, yeast hydrolysis special composite enzyme of the present invention is used for the yeast hydrolysis, and the effect of various biological enzyme coupled in common makes yeast hydrolysis liquefied ammonia base acid content greater than 0.83g/100ml, and the amino nitrogen of giving money as a gift is greater than 6.73%, and the total nitrogen of giving money as a gift is greater than 12.0%.
9, yeast hydrolysis special composite enzyme of the present invention is used for the yeast hydrolysis, make yeast hydrolyzed solution Nucleotide (I+G) content height, have very strong delicate flavour and strong sauce fragrance, good trophicity and food safety, do not need the outside Nucleotide materials such as (I+G) that adds, do not contain harmful material.
10, yeast hydrolysis special composite enzyme of the present invention is used for the yeast hydrolysis, makes yeast hydrolyate matter point of contact reasonable, nutritious, and the total nitrogen content height has very high biological culture growth property, and alternative extractum carnis is as the biological fermentation nutrition agent.
11, yeast hydrolysis special composite enzyme of the present invention, be applicable to yeast deep processings such as molasses yeast, cereuisiae fermentum, bread yeast or other process for processing waste yeasts, its yeast hydrolyzed solution can be used for aspects such as biological fermentation, seasonings, meat-based food processing, processing of aquatic products, condiment for instant noodles, puffed food and nutritional supplements.
Description of drawings:
Fig. 1 is a process flow diagram of the present invention.
Embodiment
For ease of understanding the present invention, be elaborated with specific embodiment in conjunction with the accompanying drawings, but should do not regard each embodiment as any limitation of the invention.
The preparation of yeast hydrolysis special composite enzyme, be with beta-glucanase, mannase, saccharifying enzyme, restriction endonuclease, excision enzyme, food flavor enzyme and phosphodiesterase respectively by fermentation, extraction, freeze-drying, pulverizing, mix by the optimization of C design proportion then and obtain yeast hydrolysis special composite enzyme.Concrete grammar is:
Embodiment 1
The preparation of beta-glucanase: with the soybean is raw material, pulverize, sterilization, add viride (Trichoderma viride) bacterial classification or aspergillus niger (Aspergillus niger) bacterial classification, fermented according to a conventional method 3 days, extract then, separation, ultrafiltration, freeze-drying, pulverize and obtain beta-glucanase, preserve standby.
The preparation of mannase: with the soybean is raw material, pulverize, sterilization, add viride bacterial classification (Trichoderma viride) or aspergillus niger (Aspergillus niger) bacterial classification, fermented according to a conventional method 3 days, extract then, separation, ultrafiltration, freeze-drying, pulverize and obtain mannase, preserve standby.
The preparation of saccharifying enzyme: with the soybean is raw material, pulverizes sterilization, add viride (Trichoderma viride) bacterial classification or aspergillus niger (Aspergillus niger) bacterial classification, fermented according to a conventional method 3 days, extract then, separation, ultrafiltration, freeze-drying, pulverizing obtains saccharifying enzyme, preserves standby.
The preparation of restriction endonuclease: be prepared as example with papoid, take off new fresh Chinese flowering quince syrup from pawpaw tree or fruit, earlier with tap water or distilled water or deionized water towards rare or dissolving pawpaw latex, extract by freeze thawing then, obtain papoid through micro-filtration, ultrafiltration and concentration, vacuum lyophilization, pulverizing again, sealing is preserved standby.
The preparation of excision enzyme: with the soybean is raw material, pulverize, sterilization, add aspergillus oryzae (Aspergillus oryzae) or Rhizopus oryzae (Rhizopus oryzae), aspergillus niger (Aspergillus niger) bacterial classification, 30 ℃, 90% humidity condition bottom fermentation 1 day, extraction then, micro-filtration, ultrafiltration, freeze-drying, pulverizing obtain excision enzyme, preserve standby.
The preparation of food flavor enzyme: with the soybean is raw material, pulverize, sterilization, add Mucor (Actinomucor elegans), aspergillus oryzae bacterial classifications such as (Aspergillus oryzae), 25 ℃, 70% humidity condition bottom fermentation 1 day, extraction then, micro-filtration, ultrafiltration, freeze-drying, pulverizing obtain food flavor enzyme, preserve standby.
The preparation of phosphodiesterase: with the soybean is raw material, pulverizes sterilization, add Penicillium citrinum (Penicillium citrinum) bacterial classification, 25 ℃, 70% humidity condition bottom fermentation 1 day, extraction then, micro-filtration, ultrafiltration, freeze-drying, pulverizing obtained the phosphodiesterase enzyme, preserve standby.
With above-mentioned beta-glucanase, mannase, saccharifying enzyme, restriction endonuclease, excision enzyme, food flavor enzyme and phosphodiesterase 5: 10: 17 by weight: 38: 10: 10: 10 ratio, mix, obtain yeast hydrolysis special composite enzyme.
This routine product takes ultraviolet method to carry out enzyme activity determination, wherein the beta-glucanase enzyme is lived and is 17500u/g, the mannase enzyme is lived and is 70000u/g, the enzyme of saccharifying enzyme is lived and is that it is 470000u/g that 115000u/g, the enzyme of restriction endonuclease live, and the enzyme of excision enzyme is lived and is 50000u/g, the enzyme of food flavor enzyme is lived and is 50000u/g, the enzyme of phosphodiesterase 70000u/g alive places 10 ℃ of preservations, guarantees the quality in 12 months.
Present embodiment product application result:
(1) hydrolysis molasses yeast: earlier the molasses yeast is added water and be transferred to 8～20% concentration, under the temperature of the pH value 4.5～7.0 and 45～70 ℃, add the product yeast hydrolysis special composite enzyme that present embodiment obtains by 0.1～0.6% of substrate weight, enzymolysis 10～25 hours the results are shown in Table 1.
Table 1: hydrolysis molasses yeast is table as a result
(2) hydrolysis cereuisiae fermentum: earlier cereuisiae fermentum is added water and be transferred to 8～20% concentration, under the temperature of the pH value 4.5～7.0 and 45～70 ℃, add the product yeast hydrolysis special composite enzyme that present embodiment obtains by 0.1～0.6% of substrate weight, enzymolysis 10～25 hours the results are shown in Table 2.
Table 2: the hydrolysis cereuisiae fermentum is table as a result
Embodiment 2
The preparation of beta-glucanase: with the dregs of beans is raw material, pulverize, sterilization, add viride (Trichoderma viride) bacterial classification or aspergillus niger (Aspergillus niger) bacterial classification, fermented according to a conventional method 4 days, extract then, separation, ultrafiltration, freeze-drying, pulverize and obtain beta-glucanase, preserve standby.
The preparation of mannase: with the dregs of beans is raw material, pulverize, sterilization, add viride (Trichoderma viride) bacterial classification or aspergillus niger (Aspergillus niger) bacterial classification, fermented according to a conventional method 4 days, extract then, separation, ultrafiltration, freeze-drying, pulverize and obtain mannase, preserve standby.
The preparation of saccharifying enzyme: with the dregs of beans is raw material, pulverizes sterilization, add viride (Trichoderma viride) bacterial classification or aspergillus niger (Aspergillus niger) bacterial classification, fermented according to a conventional method 4 days, extract then, separation, ultrafiltration, freeze-drying, pulverizing obtains saccharifying enzyme, preserves standby.
The preparation of restriction endonuclease: be prepared as example with neutral protease, fermented extracted obtains according to a conventional method, preserves standby.
The preparation of excision enzyme: with the dregs of beans is raw material, pulverize, sterilization, add aspergillus oryzae (Aspergillus oryzae) or Rhizopus oryzae (Rhizopus oryzae), aspergillus niger (Aspergillus niger) bacterial classification, 40 ℃, 80% humidity condition bottom fermentation 3 days, extraction then, micro-filtration, ultrafiltration, freeze-drying, pulverizing obtain excision enzyme, preserve standby.
The preparation of food flavor enzyme: with the dregs of beans is raw material, pulverize, sterilization, add Mucor (Actinomucor elegans), aspergillus oryzae bacterial classifications such as (Aspergillus oryzae), 35 ℃, 80% humidity condition bottom fermentation 3 days, extraction then, micro-filtration, ultrafiltration, freeze-drying, pulverizing obtain food flavor enzyme, preserve standby.
The preparation of phosphodiesterase: with the dregs of beans is raw material, pulverizes sterilization, add Penicillium citrinum (Penicillium citrinum) bacterial classification, 35 ℃, 80% humidity condition bottom fermentation 3 days, extraction then, micro-filtration, ultrafiltration, freeze-drying, pulverizing obtained the phosphodiesterase enzyme, preserve standby.
With above-mentioned beta-glucanase, mannase, saccharifying enzyme, restriction endonuclease, excision enzyme, food flavor enzyme and phosphodiesterase 8: 13: 18 by weight: 40: 8: 8: 5 ratio, mix, obtain yeast hydrolysis special composite enzyme.
This routine product takes ultraviolet method to carry out enzyme activity determination, wherein the beta-glucanase enzyme is lived and is 28000u/g, the mannase enzyme is lived and is 89000u/g, the enzyme of saccharifying enzyme is lived and is that it is 500000u/g that 120000u/g, the enzyme of restriction endonuclease live, and the enzyme of excision enzyme is lived and is 40000u/g, the enzyme of food flavor enzyme is lived and is 40000u/g, the enzyme of phosphodiesterase 35000u/g alive places 10 ℃ of preservations, guarantees the quality in 12 months.
This routine product application result:
(1) hydrolysis molasses yeast: earlier the molasses yeast is added water and be transferred to 8～20% concentration, under the temperature of the pH value 4.5～7.0 and 45～70 ℃, add the product yeast hydrolysis special composite enzyme that present embodiment obtains by 0.1～0.6% of substrate weight, enzymolysis 10～25 hours the results are shown in Table 3.
Table 3: hydrolysis molasses yeast is table as a result
(2) hydrolysis cereuisiae fermentum: earlier cereuisiae fermentum is added water and be transferred to 8～20% concentration, under the temperature of the pH value 4.5～7.0 and 45～70 ℃, add the product yeast hydrolysis special composite enzyme that present embodiment obtains by 0.1～0.6% of substrate weight, enzymolysis 10～25 hours the results are shown in Table 4.
Table 4: the hydrolysis cereuisiae fermentum is table as a result
Embodiment 3
The preparation of beta-glucanase: with wheat bran is raw material, pulverize, sterilization, add viride (Trichoderma viride) bacterial classification or aspergillus niger (Aspergillus niger) bacterial classification, fermented according to a conventional method 3 days, extract then, separation, ultrafiltration, freeze-drying, pulverize and obtain beta-glucanase, preserve standby.
The preparation of mannase: with wheat bran is raw material, pulverize, sterilization, add viride (Trichoderma viride) bacterial classification or aspergillus niger (Aspergillus niger) bacterial classification, fermented according to a conventional method 3 days, extract then, separation, ultrafiltration, freeze-drying, pulverize and obtain mannase, preserve standby.
The preparation of saccharifying enzyme: with wheat bran is raw material, pulverizes sterilization, add viride (Trichoderma viride) bacterial classification or aspergillus niger (Aspergillus niger) bacterial classification, fermented according to a conventional method 3 days, extract then, separation, ultrafiltration, freeze-drying, pulverizing obtains saccharifying enzyme, preserves standby.
The preparation of restriction endonuclease: be prepared as example with Sumizyme MP, fermented extracted obtains according to a conventional method, preserves standby.
The preparation of excision enzyme: with wheat bran is raw material, pulverize, sterilization, add aspergillus oryzae (Aspergillus oryzae) or Rhizopus oryzae (Rhizopus oryzae), aspergillus niger (Aspergillus niger) bacterial classification, 50 ℃, 90% humidity condition bottom fermentation 5 days, extraction then, micro-filtration, ultrafiltration, freeze-drying, pulverizing obtain excision enzyme, preserve standby.
The preparation of food flavor enzyme: with wheat bran is raw material, pulverize, sterilization, add Mucor (Actinomucor elegans), aspergillus oryzae bacterial classifications such as (Aspergillus oryzae), 45 ℃, 90% humidity condition bottom fermentation 4 days, extraction then, micro-filtration, ultrafiltration, freeze-drying, pulverizing obtain food flavor enzyme, preserve standby.
The preparation of phosphodiesterase: with wheat bran is raw material, pulverizes sterilization, add Penicillium citrinum (Penicillium citrinum) bacterial classification, 45 ℃, 90% humidity condition bottom fermentation 4 days, extraction then, micro-filtration, ultrafiltration, freeze-drying, pulverizing obtained the phosphodiesterase enzyme, preserve standby.
With above-mentioned beta-glucanase, mannase, saccharifying enzyme, restriction endonuclease, excision enzyme, food flavor enzyme and phosphodiesterase 10: 15: 20 by weight: 45: 5: 2: 3 ratio, mix, obtain yeast hydrolysis special composite enzyme.
This routine product takes ultraviolet method to carry out enzyme activity determination, wherein the beta-glucanase enzyme is lived and is 35000u/g, the mannase enzyme is lived and is 100000u/g, the enzyme of saccharifying enzyme is lived and is that it is 560000u/g that 140000u/g, the enzyme of restriction endonuclease live, and the enzyme of excision enzyme is lived and is 25000u/g, the enzyme of food flavor enzyme is lived and is 11000u/g, the enzyme of phosphodiesterase 22000u/g alive places 10 ℃ of preservations, guarantees the quality in 12 months.
This routine product application result:
(1) hydrolysis molasses yeast: earlier the molasses yeast is added water and be transferred to 8～20% concentration, under the temperature of the pH value 4.5～7.0 and 45～70 ℃, add the product yeast hydrolysis special composite enzyme that present embodiment obtains by 0.1～0.6% of substrate weight, enzymolysis 10～25 hours the results are shown in Table 5.
Table 5: hydrolysis molasses yeast is table as a result
(2) hydrolysis cereuisiae fermentum: earlier cereuisiae fermentum is added water and be transferred to 8～20% concentration, under the temperature of the pH value 4.5～7.0 and 45～70 ℃, add the product yeast hydrolysis special composite enzyme that present embodiment obtains by 0.1～0.6% of substrate weight, enzymolysis 10～25 hours the results are shown in Table 6.
Table 6: the hydrolysis cereuisiae fermentum is table as a result
Claims (10)
1. yeast hydrolysis special composite enzyme, it is characterized in that: comprise beta-glucanase, mannase, saccharifying enzyme, restriction endonuclease, excision enzyme, food flavor enzyme and phosphodiesterase, its composition and parts by weight thereof are: beta-glucanase is 1～30 part, mannase is 1～30 part, saccharifying enzyme is 1～30 part, and restriction endonuclease is 15～65 parts, and excision enzyme is 1～10 part, food flavor enzyme is 1～10 part, and phosphodiesterase is 1～10 part.
2. the preparation method of yeast hydrolysis special composite enzyme according to claim 1, it is characterized in that: earlier with beta-glucanase, mannase, saccharifying enzyme, restriction endonuclease, excision enzyme, food flavor enzyme and phosphodiesterase respectively by fermentation, extraction, freeze-drying, pulverizing, evenly obtain yeast hydrolysis special composite enzyme by the formulating of recipe mixed then, its technological process is:
(1) preparation of beta-glucanase: with soybean or dregs of beans or wheat bran is raw material, adopts viride (Trichoderma viride) bacterial classification or aspergillus niger (Aspergillus niger) bacterial classification to prepare according to a conventional method, preserves standby;
(2) preparation of mannase: with soybean or dregs of beans or wheat bran is raw material, adopts viride (Trichoderma viride) bacterial classification or aspergillus niger (Aspergillus niger) bacterial classification to prepare according to a conventional method, preserves standby;
(3) preparation of saccharifying enzyme: with soybean or dregs of beans or wheat bran is raw material, adopts viride (Trichoderma viride) bacterial classification or aspergillus niger (Aspergillus niger) bacterial classification to prepare according to a conventional method, preserves standby;
(4) preparation of restriction endonuclease: taking off new fresh Chinese flowering quince syrup with pawpaw tree or fruit is raw material, earlier with tap water or distilled water or deionized water towards rare or dissolving pawpaw latex, extract by freeze thawing then, obtain papoid through micro-filtration, ultrafiltration and concentration, vacuum lyophilization, pulverizing again, preserve standby; The preparation of neutral protease, Sumizyme MP obtains through fermented extracted according to a conventional method;
(5) preparation of excision enzyme, with soybean or dregs of beans or wheat bran is raw material, pulverize, sterilization, add aspergillus oryzae (Aspergillus oryzae) or Rhizopus oryzae (Rhizopus oryzae), aspergillus niger (Aspergillus niger) bacterial classification, 30～50 ℃, 70～90% humidity condition bottom fermentation 1～5 day, extraction then, micro-filtration, ultrafiltration, freeze-drying, pulverizing obtained, and preserve standby;
(6) preparation of food flavor enzyme, with soybean or dregs of beans or wheat bran is raw material, pulverize, sterilization, add Mucor (Actinomucor elegans), aspergillus oryzae (Aspergillus oryzae) bacterial classification, 25～45 ℃, 70～90% humidity condition bottom fermentation 1～4 day, extraction then, micro-filtration, ultrafiltration, freeze-drying, pulverizing obtained, and preserve standby;
(7) preparation of phosphodiesterase, with soybean or dregs of beans or wheat bran is raw material, pulverize, sterilization, add Penicillium citrinum (Penicillium citrinum) bacterial classification, 25～45 ℃, 70～90% humidity condition bottom fermentation 1～4 day, extraction then, micro-filtration, ultrafiltration, freeze-drying, pulverizing obtained, and preserve standby;
(8), mix by formulating of recipe composition and parts by weight thereof and to obtain yeast hydrolysis special composite enzyme with the above-mentioned beta-glucanase for preparing, mannase, saccharifying enzyme, restriction endonuclease, excision enzyme, food flavor enzyme and phosphodiesterase.
3. yeast hydrolysis special composite enzyme according to claim 1, it is characterized in that: described beta-glucanase, mannase, saccharifying enzyme, be to extract by viride bacterial classification or aspergillus niger strain fermentative preparation to obtain, wherein the beta-glucanase enzyme activity is 1000～100000u/g, the mannase enzyme activity is 5000～200000u/g, and the glucoamylase enzyme vigor is 5000～200000u/g.
4. yeast hydrolysis special composite enzyme according to claim 1, it is characterized in that: described restriction endonuclease, the mixture that comprises papain, neutral protease and Sumizyme MP, blending ratio are 0～5:0～5:0～5, and its total enzyme activity is 10000～800000u/g.
5. yeast hydrolysis special composite enzyme according to claim 1 is characterized in that: described excision enzyme, comprise the biological enzyme that one or more strain fermentations in aspergillus oryzae or Rhizopus oryzae, the aspergillus niger prepare, and its enzyme activity is 1000～50000u/g.
6. yeast hydrolysis special composite enzyme according to claim 1 is characterized in that: described food flavor enzyme, comprise the biological enzyme that one or both strain fermentations in Mucor, the aspergillus oryzae prepare, and its enzyme activity is 1000～50000u/g.
7. yeast hydrolysis special composite enzyme according to claim 1 is characterized in that: described phosphodiesterase is to extract the phosphodiesterase that obtains from the Penicillium citrinum fermented product, and its enzyme activity is 1000～70000u/g.
8. the application method of yeast hydrolysis special composite enzyme according to claim 1, it is characterized in that: waste yeast is added water be transferred to 8～20% concentration, under the temperature of the pH value 4.5～7.0 and 45～70 ℃, press 0.1～0.6% of substrate weight and add yeast hydrolysis special composite enzyme, enzymolysis 10～25 hours, be warmed up to then more than 85 ℃ and continue 15 minutes～1 hour the hydrolyzed solution enzyme that goes out, detect the hydrolyzed solution aminoacids content greater than 0.83g/100ml, give money as a gift amino nitrogen greater than 6.73%, give money as a gift total nitrogen greater than 12.0%, separate again, concentrate, drying obtains yeast extract product at last.
9. the application of yeast hydrolysis special composite enzyme as claimed in claim 1 aspect molasses yeast, cereuisiae fermentum, bread yeast or other process for processing waste yeast yeast deep processings.
10. the application of yeast hydrolysis special composite enzyme as claimed in claim 1 aspect biological fermentation, seasonings, meat-based food processing, processing of aquatic products, condiment for instant noodles, puffed food and nutritional supplements.
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| WO2017050629A1 (en) * | 2015-09-21 | 2017-03-30 | Dsm Ip Assets B.V. | Yeast cell wall derived flavour |
| CN105950481B (en) * | 2016-05-30 | 2019-05-03 | 湖北工业大学 | One Aspergillus oryzae bacterial strain and its protease of generation are produced applied to yeast extract |
| CN106520875A (en) * | 2016-11-21 | 2017-03-22 | 安徽省华信生物药业股份有限公司 | Process for preparing selenium yeast protein peptide powder from selenium-rich yeast |
| CN109198156B (en) * | 2017-06-30 | 2022-03-08 | 安琪纽特股份有限公司 | Yeast protein and preparation method and application thereof |
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| CN107594451A (en) * | 2017-08-30 | 2018-01-19 | 珠海天香苑生物科技发展股份有限公司 | The method that aspergillus oryzae prepares yeast extract |
| CN107853706A (en) * | 2017-10-13 | 2018-03-30 | 浙江东成生物科技股份有限公司 | A kind of preparation method of acidproof salt-resistant type yeast extract |
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