CN104357337A - Rhizopus for food fermentation and application - Google Patents
Rhizopus for food fermentation and application Download PDFInfo
- Publication number
- CN104357337A CN104357337A CN201410676014.5A CN201410676014A CN104357337A CN 104357337 A CN104357337 A CN 104357337A CN 201410676014 A CN201410676014 A CN 201410676014A CN 104357337 A CN104357337 A CN 104357337A
- Authority
- CN
- China
- Prior art keywords
- zsm
- rhizopus
- rhizopus oryzae
- cctcc
- fermentation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- 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
- C12R2001/845—Rhizopus
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/02—Preparation of other alcoholic beverages by fermentation
-
- 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
-
- 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
- C12N1/16—Yeasts; Culture media therefor
Abstract
The invention belongs to the field of food fermentation, and in particular relates to rhizopus for food fermentation and application. Rhizopus oryzae zsm-003, zsm-004 and rhizopus zsm-005 suitable for rice product and sweet rice wine fermentation are separated from sweet rice wine. A microorganism complex bacterium agent for food fermentation is developed, which comprises rhizopus oryzae zsm-003, zsm-004 and brettanomyces custersii ZSM-001. Strain survival rate of the complex fermenting agent reaches more than 35%, and the produced sweet rice wine is relatively rich in sugar composition, intense in flavor, good in taste, and relatively high in contents of delicious amino acid and essential amino acid. Furthermore, the invention discloses a preparation method and application of the complex fermenting agent.
Description
Technical field
The invention belongs to food fermentation and food processing technology field, be specifically related to a kind of head mold of food fermentation and application, the present invention includes the bacterial screening of microbial starter culture, and utilize many bacterial strains prepare compound ferment and the application in rice food fermentative production thereof.
Background technology
Head mold (Rhizopus) belongs to the Rhizopus (Rhizopus Ehrenberg) in the mucorales (Mucorales) of Eucaryotae, Mucoraceae (Mucoraceae) on taxonomy.It is of a great variety, wherein outstanding with amylase that head mold produces enzyme, is a kind of very important brewing microorganism.Head mold can produce a large amount of amylase in its growth metabolism process, and has very strong activity, amylan can be resolved into fermentable sugar thoroughly.In addition, head mold can also produce a series of mould systems such as certain proteolytic enzyme, zymase and lipase, and produces the organic acids such as citric acid, lactic acid and succsinic acid at fermenting process, has important effect in wine industry.Along with the development of modern biotechnology, people are to the research of head mold and utilize further deeply.The research of head mold is not only confined to liquor industry, and applies to various healthcare products, lactic acid, zymin gradually.In addition, people also utilize the adsorptive power of some head mold heavy metal and are applied to environmental improvement.
Sweet wine take glutinous rice as the fermentation rice made products that main raw material is brewageed through boiling, inoculation mixed song, microorganism diastatic fermentation.Glutinous rice is through boiling gelatinization, under the acting in conjunction of the meta-bolitess such as microorganism and enzyme thereof such as the mould in distiller's yeast, the macromolecular substance such as starch, protein, fat are degraded to nutrition and the flavor characteristic that the small-molecule substances such as the monose such as glucose, oligose, peptide class, amino acid, organic acid give sweet wine uniqueness, contribute to improving immunizing power, enhancing metabolism, and the effect having relaxing the muscles and stimulate the blood circulation, promote longevity, be used as a kind of nourishing food by people.Tradition sweet wine starter is generally that access seed microbial growth post-drying is shaping, mainly contains the three major types microorganisms such as yeast, bacterium, mould with grain (based on waxy rice) and Chinese herbal medicine powder for raw material.In distiller's yeast, each quasi-microorganism is of a great variety, and most common microbiological has yeast saccharomyces cerevisiae, head mold, aspergillus oryzae, Mucor etc., has the several functions such as saccharification, fermentation, esterification Sum decomposition protein.At the brewing process of sweet wine, nutritive substance in raw material decomposes and metabolism by microorganism, form sugar, acid, alcohols and the various meta-bolites such as amino acid, peptide class, these materials form fragrance matter or its fragrance precursor material and the nutritive substance of sweet wine, determine quality of finished.
Handicraft workshop formula sweet wine starter among the people forms jointly primarily of the multiple-microorganisms such as head mold, Mucor, yeast and the herbal medicine such as ground rice, Herba polygoni hydropiperis, and the sweet wine local flavor brewageed is mellow, mouthfeel is soft.And modern throw type leaven is by known head mold and yeast or single pure head mold adds protective material and stopping composition is made.Distiller's yeast among the people causes sweet wine local flavor there are differences because the complete processing of its auxiliary material, stopping composition and region is different, and simultaneously during the fermentation because the activity of bacterial classification and ratio easily change, the quality stability of its product is poor.And in direct-throwing microbial starter culture bacterial activity and ratio more stable, microorganism concn is high, and inoculum size is little, can shorten fermentation time, and effectively prevents varied bacteria growing, thus ensures the stable of quality product.Meanwhile, direct-throwing microbial starter culture can be developed to special leaven for the characteristic of product.The defects such as the starter quality of suitability for industrialized production is more stable, but the sweet wine that fermentation obtains there is local flavor and mouthfeel is single, and vinosity sense is not enough.Adopt fermented by mixed bacterium can improve industrial fermentation agent to a certain extent for the production of sweet wine and make the thin shortcoming of sweet wine local flavor.
Important procedure in solid fermentation agent is prepared in drying.Conventional fluidised bed drying speed is fast, but easily causes the pollution of bacterial classification.Bacterium powder vigor prepared by vacuum lyophilization is high, but equipment power consumption is large.And though spraying dry, warm air drying efficiency are high, the survival rate of thalline is lower.The microwave drying of rising in recent years have dehydration rapidly, be heated comparatively evenly, the feature such as environmental protection.
At present, sweet wine special leaven production technology is ripe gradually.Such as publication number is the patent application of CN 102604840 A, and disclose the rice wine koji and preparation method that utilize the rhizopus filtered out to prepare, the sweet wine brewageed with it can ensure alcoholic strength and the pol of higher level.Publication number is the patent application of CN 102994396 A, disclose with pure song and mixed starters by a certain percentage mixing manufacture rice wine koji improve the local flavor of rice wine.Xiaogan sweet wine have delicate fragrance attack people, sweet tasty and refreshing, dense and be not stained with, rare and unique flavor characteristic such as not flow, be " China's time-honored brand " green food.In its traditional distiller's yeast, microorganism is formed the flavor characteristic of Xiaogan sweet wine uniqueness and has important effect.Due to microbic activity and the meta-bolites difference of different sources in distiller's yeast, the local flavor of the sweet wine product of its brew and nutrition is caused to differ from one another.Some bacterial strain can produce characteristic metabolic products, and its product has higher nutritive value to a certain extent.
Traditional characteristics rice wine fermenting microorganism is indefinite, and the product quality feature of making is unclear, and is difficult to industrialization; And pure strain starter causes the single grade of product special flavour to be all this area technical problem urgently to be resolved hurrily.
Summary of the invention
The object of the invention is to the defect overcoming prior art, there is provided a kind of head mold of food fermentation screening and application, the present invention includes the bacterial screening of microbial starter culture, utilize many bacterial strains such as the yeast of rhizopus and screening, adopt the compound ferment being prepared into applicable sweet wine fermentative production of microwave drying technology.
Realizing method of the present invention is, from Xiaogan Prefecture of Hubei Province Traditional Folk koji, to obtain three strain leavening properties good for screening and separating, these bacterial strains belong to rhizopus and yeast, above-mentioned three strain bacterial strains are made compound microbial culture starter, rice food particularly sweet wine nutritive value can be significantly improved, adapt to industrial fermentation technique.
The present invention is realized by following technical proposal:
Applicant is by being separated from Hubei Province's Xiaogan City Traditional Folk koji, and therefrom screening obtains three strains and is applicable to food fermentation and is particularly applicable to rice food, the rhizopus strains of such as particularly sweet wine fermentative production, these bacterial strains respectively:
The Rhizopus oryzae of one strain food fermentation, be named as Rhizopus oryzae zsm-003, China is delivered on August 8th, 2014. Wuhan. Wuhan University's China typical culture collection center (CCTCC) preservation, preserving number is CCTCC NO:M2014374;
The Rhizopus oryzae of one strain food fermentation, be named as Rhizopus oryzae zsm-004, China is delivered on August 8th, 2014. Wuhan. Wuhan University's China typical culture collection center (CCTCC) preservation, preserving number is CCTCC NO:M2014375;
The head mold of one strain food fermentation, is named as Rhizopus sp.zsm-005, delivers China on August 8th, 2014. Wuhan. and Wuhan University's China typical culture collection center (CCTCC) preservation, its preserving number is CCTCC NO:M2014376.
Applicant, based on above-mentioned three strain bacterial strains, develops a kind of complex microbial inoculum of food fermentation, and this microbial composite starter includes the Rhizopus oryzae Rhizopus oryzae zsm-003 that preserving number is CCTCC NO:M 2014374; Preserving number is the Rhizopus oryzae Rhizopus oryzae zsm-004 of CCTCC NO:M 2014375; Karst Brettanomyces (Brettanomyces custersii) ZSM-001 of preserving number to be the head mold Rhizopus sp.zsm-005 of CCTCC NO:M2014376 and preserving number be CCTCC NO:M207150, the patent No. is ZL2007100536112.
Through application test, mentioned microorganism compound ferment has features:
1) in this compound ferment, the survival rate of bacterial classification is higher, reaches more than 35%;
2) this compound ferment produce sweet wine juice in carbohydrate composition abundanter, product flavour is strong, rich in taste, and containing fructose and the function such as maltose, trisaccharide maltose oligose, nutritive value is higher;
3) have higher delicious amino acid in the sweet wine product that this compound ferment is produced, necessary amino acid ratio is higher.
The separation of bacterial strain of the present invention, screening and identification:
Techniqueflow of the present invention is shown in Fig. 1.
Rhizopus oryzae Rhizopus oryzae zsm-003 (CCTCC NO:M 2014374) of the present invention, Rhizopus oryzaezsm-004 (CCTCC NO:M 2014375), the separation of Rhizopus sp.zsm-005 (CCTCC NO:M 2014376), screening and identification, with reference to Zhou Deqing chief editor, " Microbiology Experiment handbook ", Shanghai science tech publishing house, 1986; Hu Ruiqing translates, " common and conventional fungi ", Science Press, 1973.
The mycology feature of isolated strains:
The colony morphology characteristic of Rhizopus oryzae of the present invention (Rhizopus oryzae) zsm-003 (CCTCC NO:M2014374), Rhizopus oryzae (Rhizopusoryzae) zsm-004 (CCTCC M 2014375) and head mold (Rhizopus sp.) zsm-005 (CCTCC M 2014376) bacterial strain is as follows:
Rhizopus oryzae (Rhizopus oryzae) zsm-003, after potato culture cultivates 12h, colony diameter is about 1.5 ~ 2cm, and bacterium colony is more loose, and aerial hyphae is longer, and edge mycelia is more neat.Be paved with flat board after 20h, colony edge is divergent shape, occurs white and black spore.
Rhizopus oryzae (Rhizopus oryzae) zsm-004, after potato culture cultivates 12h, colony diameter is about 0.8 ~ 1.2cm, and flat plate bottom has relatively fine and close one deck mycelia, and edge mycelia is more neat.Be paved with flat board after 20h, occur white and black spore.
Head mold (Rhizopus sp.) zsm-005, after potato culture cultivates 12h, colony diameter is about 1 ~ 1.5cm, and mycelia is finer and close, and aerial hyphae growth is slower.Be paved with flat board after 24h, occur white and black spore.
The microcosmic morphological feature following (see Fig. 2) of Rhizopus oryzae of the present invention (Rhizopus oryzae) zsm-003 (CCTCC NO:M2014374), Rhizopus oryzae (Rhizopusoryzae) zsm-004 (CCTCC M 2014375) and head mold (Rhizopus sp.) zsm-005 (CCTCC M 2014376) bacterial strain:
Rhizopus oryzae (Rhizopus oryzae) zsm-003 and Rhizopus oryzae (Rhizopus oryzae) zsm-004, rhizoid is more flourishing, sporangiophore vertical growth, the spherical or almost spherical of sporocyst, brown (diameter 90 ~ 120um).Spherical or the almost spherical of columella, sporangiospore is spherical, has chlamydospore.
Head mold (Rhizopus sp.) zsm-005, have rhizoid, sporangiophore vertical growth, sporocyst is subsphaeroidal, beige, diameter 50 ~ 200um, and sporangiospore is spherical or oval, has chlamydospore.
Identify two Rhizopus oryzae bacterial strains of above-mentioned separation screening and the 18SrDNA sequence of a strain rhizopus strains, concrete steps are as follows:
With fungi universal primer (P1 (5'-GTAGTCATATGCTTGTCTC-3 '), P2 (5 '-CTTCCGTCAATTCCTTTAAG-3 ')) the isolated strains Rhizopus oryzaezsm-003 that obtains for sequencing primer, Rhizopus oryzae zsm-004 and Rhizopus sp.zsm-005 effective dna sequence, the 18S rDNA ordered sequence recorded carries out Blast retrieval in ncbi database.Result for retrieval is bacterial strain Rhizopus oryzae zsm-003, the sequence of Rhizopusoryzae zsm-004 and Rhizopus sp.zsm-005 respectively with Rhizopus oryzae (Rhizopus oryzac), head mold (Rhizopussp.) sequence similarity, homology is up to 99%, the sequence larger with this sequence similarity is loaded in MEGA4 software package, carry out multiple sequence comparative analysis, phylogenetic tree construction, obtains result as shown in Figure 3.
The present invention, compared with existing sweet wine fermentation strain, has the following advantages:
(1) this strain growth speed, produces black spore at fermenting process less.
(2) this bacterial strain can as high-quality special leaven: the sweet wine juice Determination of Free Amino Acids of starter brew of the present invention is higher, containing functional oligoses etc. such as maltose, carbohydrate composition abundant (embodiment 3), nutritive value is higher, and constant product quality, excellent flavor.
(3) compound ferment of the present invention achieves the complex ferment of mixed strains Karst Brettanomyces (Brettanomyces custersii) ZSM-001 and Rhizopus oryzae, compare single root arrhizus fermentation sweet wine, its fragrance is stronger, and flavour has more mellow sense.Use microwave drying process to prepare starter, there is rate of drying fast, easy to control, consume energy little, and the survival rate of bacterial strain is higher.
Accompanying drawing explanation
Sequence table SEQ ID NO:1 is the 18SrDNA nucleotide sequence of Rhizopus oryzae (Rhizopus oryzae) the zsm-003 bacterial strain that the present invention is separated.
Sequence table SEQ ID NO:2 is the 18SrDNA nucleotide sequence of Rhizopus oryzae of the present invention (Rhizopus oryzae) zsm-004 bacterial strain.
Sequence table SEQ ID NO:3 is the 18SrDNA nucleotide sequence of head mold of the present invention (Rhizopus sp.) zsm-005.
Fig. 1: the techniqueflow chart being being separated of three bacterial strain zsm-003, zsm-004 with zsm-005 of the present invention, screening and qualification.
Fig. 2: the microscopic pattern figure being three bacterial strain zsm-003, zsm-004 and zsm-005 of the present invention.Description of symbols in figure: Fig. 2 a is that (a figure in Fig. 2 a is rhizoid to Rhizopus oryzae zsm-003 bacterial strain microscopic pattern figure; B figure in Fig. 2 a is sporocyst; C figure in Fig. 2 a is sporangiospore; D figure in Fig. 2 a is chlamydospore).
Fig. 2 b is that (a figure in Fig. 2 b is rhizoid to Rhizopus oryzae zsm-004 bacterial strain microscopic pattern figure; B figure in Fig. 2 b is chlamydospore; C figure in Fig. 2 b is the sporocyst of division; D figure in Fig. 2 b is intact spore capsule).
Fig. 2 c is that (a figure in Fig. 2 c is rhizoid to head mold zsm-005 bacterial strain microscopic pattern figure; B figure in Fig. 2 c is sporocyst; C figure in Fig. 2 c is sporangiospore; D figure in Fig. 2 c is chlamydospore).
Fig. 3: the evolution tree graph being three bacterial strain zsm-003, zsm-004 and zsm-005 of the present invention.Wherein: Fig. 3 a is the analysis of zsm-003 phylogenetic tree; Fig. 3 b is the phylogenetic tree analysis of zsm-004; Fig. 3 c is the phylogenetic tree analysis of zsm-005.
Fig. 4: the techniqueflow chart being solid-state composite fermenting agent of the present invention and single fermentation agent.
Embodiment
Embodiment 1: the separation screening of aimed strain and qualification
1, separation method (dilution-plate method)
Under aseptic technique field, get 10.0g bent from the folk tradition sweet wine of Hubei Province's Xiaogan City, join in the triangular flask of 90ml sterilized water and (in bottle, put the little granulated glass sphere of some amount in advance), 30min is rocked in vibration, then carry out gradient dilution, select extent of dilution to be the diluent of 10-3,10-4,10-5.Get 1ml under aseptic condition and dilute sample liquid, be transferred in culture dish, then pour PDA substratum into, cultivate at 30 DEG C after mixing.Picking plane Superior fungus mycelium is started after 12h, respectively dibbling in Cha Shi substratum (Li Jianrong, Cai Aiqun. the Isolation and Identification [J] of micro-ox thing wanted by folk tradition distiller's yeast card. brewing science and technology, 2007, l:111-121.), 48h is cultivated for 30 DEG C.After dull and stereotyped purifying four generation, turn PDA inclined-plane (for conventional substratum) storage.
2, authentication method
The morphological feature of 2.1 bacterial strains edits " Microbiology Experiment handbook " with reference to Zhou Deqing, " common and conventional fungi ", Science Press, the method introduced for 1973.
2.2 utilize inserted sheet method to observe mould microscopic pattern: by sterile cover slips with 45
.the angle of left and right is inserted in PDA plate culture medium side by side, and depth of penetration is about 1/3 of cover glass.With the boundary line place of aseptic inoculation ring picking fungal hyphae dibbling in cover glass and substratum, cultivate 24h at 30 DEG C.With tweezers, cover glass is carefully taken out, clean substratum and the mycelia at the cover glass back side, be placed on slide glass and (have one of mycelia to face down), directly observe under being placed in microscope.
2.3 biomolecular science qualifications:
(1) the genomic extraction of head mold: transfer in the PDA liquid nutrient medium of 50mL after strains tested is activated, 30 DEG C of shaking table (200r/min) overnight incubation.Collected by centrifugation mycelium, and employing CTAB method (Xu Lu. the application .2007. of molecular marking technique in Monascus anka Nakazawa et sato bacterium identification) extract genomic dna, detect extracting genome DNA quality through 0.8% agarose gel electrophoresis.If up-to-standard ,-20 DEG C save backup.
(2) pcr amplification: adopt fungi l 8SrDNA universal primer: P1 (5'-GTAGTCATATGCTTGTCTC-3'), P2 (5'-CTTCCGTCAATTCCTTTAAG-3') with the DNA genome of head mold for template, the partial gene sequence (its nucleotide sequence is as shown in sequence table SEQ ID NO:1) of pcr amplification head mold 18S rDNA.Pcr amplification condition: 94 DEG C of denaturation 5min; 94 DEG C of sex change 30s, 53 DEG C of annealing 30s, 72 DEG C extend 2min, 30 circulations; 72 DEG C extend 7min.The PCR thing getting 2uL carries out electrophoresis detection in 0.8% sepharose.
(9) pcr amplification product after 0.8% agarose gel electrophoresis inspection is carried out 18SrDNA sequential analysis, sequencing is completed by Nanjing Jin Sirui Bioisystech Co., Ltd.In ncbi database, adopted by sequencing result Blast software to carry out online compare of analysis, and by MEGA4 software building evolutionary tree.Determine that bacterial strain ZSM-003, ZSM-004 of the present invention be Rhizopus oryzae (Rhizopus oryzae), ZSM-005 are head mold (Rhizopus sp.).
Embodiment 2 utilizes bacterial strain production microbial composite starter of the present invention
1, experiment material
Karst Brettanomyces (Brettanomyces custersii) the ZSM-001 preserving number that the bacterial strain tested make use of the former isolation identification of applicant is CCTCC NO:M207150, the patent No. is ZL2007100536112) and Rhizopus oryzae Rhizopusoryzae zsm-003 (CCTCC NO:M 2014374), Rhizopus oryzae zsm-004 (CCTCC NO:M 2014375), Rhizopus sp.zsm-005 (CCTCC NO:M 2014376).
2, the making of solid-state single culture starter and solid-state microorganism composite fungus agent
1) cultivation of test tube strains: by Karst Brettanomyces ZSM-001 and Rhizopus oryzae Rhizopus oryzae zsm-003 (CCTCCNO:M 2014374), Rhizopus oryzae zsm-004 (CCTCC NO:M 2014375), Rhizopus sp.zsm-005 (CCTCC NO:M 2014376) is inoculated in inclined-plane PDA substratum, under 30 DEG C of conditions, cultivate 2 ~ 3d, make it activate and be test tube slant bacterial classification;
2) triangular flask enlarged culturing: by step 1) in the triangular flask potato liquid nutrient medium of inoculation after sterilizing in (in 1L distilled water, add the fresh potato of 200g peeling, boiling water bath 30min, filtered through gauze, moisturizing is to 1L), under 30 DEG C of conditions, cultivate 24h, make its thalline viable count reach 10
6~ 10
7cfu/ml, for subsequent use.
3) bacterium liquid mixing: by step 2) in Karst Brettanomyces ZSM-001 bacterium liquid and Rhizopus oryzaezsm-003 (CCTCC NO:, 1) in, 1) M 2014374 in), Rhizopus oryzae zsm-004 (CCTCC NO:M 2014375), Rhizopus sp.zsm-005 (CCTCC NO:M 2014376)) be single activated rhizopus suspension in 2:3:3:2 ratio (ml) mixing (or 1) by volume) add 7.5 ~ 10% trehalose and 3 ~ 5% glycerine as protective material, obtain mixed bacteria liquid;
4) dry: by step 3) in mixed bacteria liquid be in mass ratio 8.5% ~ 10% ratio add stopping composition (such as dry rice flour), mixing, WL2S-1 type microwave oven (the happy new and high technology engineering corporation in Nanjing three), microwave dose is 1w/g, dry 1.5min, interval 0.5min, processes 2 ~ 5 times repeatedly, until water content is about 6%, namely obtain solid-state single or compound ferment.
5) (10 are about with the solid-state single microbial inoculum described in polyethylene packaging bag hermetic package
7~ 10
8cfu/g) or compound ferment (bacteria containing amount is about 10
7~ 10
8cfu/g), 4 DEG C of refrigerator storages are placed in for subsequent use.
The solid fermentation agent made is carried out to the detection of the bacterial classification survival rate of different dry drying method, result shows to adopt the survival rate (38.11 ± 0.88%) of bacterial classification in the drying solid-state single microbial inoculum of method for microwave drying or microbial composite starter to be significantly higher than traditional warm air drying (25.67 ± 1.01%), and the time of microwave drying shortlyer can enhance productivity greatly.Note: cell viable count adopts the method for plate count to obtain, calculates director and is:
Rate of loss (%)=supernatant liquor viable count/initial viable count × 100%;
Survival rate (%)=sedimentation cell viable count/(initial viable count-supernatant liquor viable count) × 100%; Significant difference (p<0.05) is there is between a, b letter representation column data.
The application of embodiment 1 solid fermentation agent of the present invention in sweet wine fermentative production
Single solid fermentation microbial inoculum and composite solid starter make the sensory evaluation scores of sweet wine
The sweet wine sensory evaluation scores that the different single bacterial strain of table 1 makes
Note: zsm-003 represents the single solid fermentation agent made of zsm-003; Zsm-004 represents the single solid fermentation agent made of zsm-004; Zsm-005 represents the single solid fermentation agent made of zsm-005; FP represents the composite solid starter with the composite making of three strain bacterium.The sexy official's evaluation assessment of quantitative description, chooses 10 systems, 0-9 (0=is insipid, and 9=taste is the strongest).
As shown in Table 1, in the sweet wine that single solid fermentation microbial inoculum makes, the product tart flavour of application zsm-003 bacterial strain is more outstanding, the sour-sweet sense of product of zsm-005 bacterial strain is softer, the product integrated sensory of zsm-004 bacterial strain marks better, and utilize zsm-003 bacterial strain, the sensory evaluation scores of compound ferment prepared by zsm-005 and zsm-004 tri-bacterial strain is the highest.In table 1.
1, the sensory evaluation of the sweet wine juice of different fermentations microbial inoculum making
Sensory evaluation method reference: Zhu Haiquan. the research [D] of the exploitation of mixing koji and sweet wine production technology. Changsha: Sino-South African Forestry University of Science and Technology, the methods of 2012 reports.Result is as table 2.
The agent of table 2 different fermentations make sweet wine sensory evaluation scores (n=2,
)
Note: AQ is commercially available pure head mold distiller's yeast, and SM is commercially available traditional distiller's yeast, and FP is the solid union microbial starter culture that the present invention prepares.Same column data small English alphabet difference represents that data exist significant difference (p<0.05).
As shown in Table 2, the sweet wine that solid compound ferment of the present invention makes, on color and luster and sense organ overall score, is obviously better than the sweet wine made of pure head mold and traditional distiller's yeast.
2, the measuring method of sweet wine juice Free Amino Acids adopts National Standard of the People's Republic of China, and the standard method of standard No. GB/T5009.124-2003 measures
The sweet wine juice Free Amino Acids composition measuring that different fermentations microbial inoculum makes the results are shown in Table 3.
Table 4 sweet wine juice Free Amino Acids composition (n=2,
)
Note: in table, AQ is commercially available pure head mold distiller's yeast, and SM is commercially available traditional distiller's yeast, FP is the solid-state multiple and microbial starter culture (i.e. koji of the present invention) that the present invention prepares; 1,2,3 represent Fresh ear field, sweet taste amino acid, bitter taste amino acid respectively;
* indispensable amino acid is represented; Represent that data exist significant difference (p<0.05) with a line English alphabet difference.
As shown in Table 3, the sweet wine that makes of different distiller's yeast (or starter) under the same conditions, its aminoacids content has certain difference.In the sweet wine that the solid-state multiple and microbial starter culture that the present invention produces makes, delicious amino acid total content at most (98.56mg/100ml), and its flavour is strong.The content of sweet wine Free Amino Acids produced with solid union microbial starter culture of the present invention and the content of indispensable amino acid are all greater than other two kinds, show that nutritive value is better.
3, in the sweet wine wine juice that makes of solid compound ferment and commercial pure root arrhizus fermentation agent and commercial traditional distiller's yeast, carbohydrate ratio of components is comparatively.
In sweet wine juice, the measuring method of monose, oligose is with reference to National Standard of the People's Republic of China, and standard No. GB/T22221-2008 method detects.
In sweet wine juice, glucose, fructose, maltose, trisaccharide maltose content detection the results are shown in Table 4.
Monose in table 4 sweet wine juice, oligose composition (n=2,
)
Note: AQ is commercially available pure head mold distiller's yeast, and SM is commercially available traditional distiller's yeast, and FP is solid union microbial starter culture (or being called that composite sweet wine is bent) prepared by the present invention.Lowercase difference represents that between same column, data exist significant difference.(p<0.05)。
As shown in Table 4, compared with the sweet wine made with commercially available pure head mold and commercially available traditional distiller's yeast, the most species of the monose that solid-state composite fermenting agent fermentation of the present invention produces and oligose, containing glucose, fructose, maltose, trisaccharide maltose, its flavour is more plentiful, mellow in taste.Detect that trisaccharide maltose etc. has the oligose of certain nourishing function in the sweet wine juice only made at solid-state composite fermenting agent of the present invention, illustrate that the sweet wine that solid-state composite fermenting agent of the present invention makes has better nutritive value.
The application of embodiment 4 solid compound ferment of the present invention in fermented rice cake
1) rice material: early rice 1Kg (commercially available), cleans according to a conventional method;
2) soak: add water 2000mL, soak 20 ~ 24h in 30 DEG C;
3) preparation of Rice & peanut milk: add 1200ml water mill slurry after cleaning several times with tap water;
4) inoculate: be 3% in mass ratio, add compound microbial culture starter prepared by the present invention, make it mix with Rice & peanut milk;
5) ferment: postvaccinal Rice & peanut milk is put into 35 DEG C of incubator fermentation 5h;
6) seasoning: by step 5 after fermenting) Rice & peanut milk add the seasoning of 200g sucrose, mixing;
7) boiling: by step 6) Rice & peanut milk put into steamer, after upper steam 15min take out, namely obtain rice of the present invention
Steamed sponge cake.
Table 5 compound ferment of the present invention compares with the sense organ of the fermented rice cake that single barms starter makes, and (every full marks are 10 points, the little kingfisher of concrete grammar reference Liu. the research and development [D] of fermented rice cake starter and composite powder. Hubei: Hua Zhong Agriculture University, 2008)
Note: Y represents commercially available saccharomycetes to make fermentation agent, and F represents compound ferment of the present invention.Lowercase difference represents that between same column, data exist significant difference (p<0.05).
As shown in Table 5, compared with the fermented rice cake made with single saccharomycetes to make fermentation agent, solid compound ferment of the present invention is in conditions, and the fermented rice cake of making, at smell, flavour and comprehensive grading is all obviously better than the fermented rice cake that single culture starter makes
The matter structure feature of the fermented rice cake that table 6 compound ferment of the present invention and single barms starter make
Note: Y represents commercially available saccharomycetes to make fermentation agent, and F represents compound ferment of the present invention.Lowercase difference represents that between same column, data exist significant difference (p<0.05).
Reference, Liu little Cui, Hu Jian, Zhao Siming etc. the matter structure feature of fermented rice cake and mathematical model [J]. Chinese grain and oil journal, 2009,24 (3): 7-10.
As shown in Table 6, the fermented rice cake hardness that the fermented rice cake that commercially available yeast fermentation agent makes makes than compound ferment of the present invention is large, and the fermented rice cake that the compound ferment utilizing the present invention to prepare makes has good chewiness and rebound resilience.
Claims (10)
1. Rhizopus oryzae (Rhizopus oryzae) zsm-003 of a strain food fermentation, be deposited in China typical culture collection center, its preserving number is CCTCC NO:M 2014374.
2. Rhizopus oryzae (Rhizopus oryzae) zsm-004 of a strain food fermentation, be deposited in China typical culture collection center, its preserving number is CCTCC NO:M 2014375.
3. head mold (Rhizopus sp.) zsm-005 of a strain food fermentation, be deposited in China typical culture collection center, its preserving number is CCTCC NO:M 2014376.
4. a microbial composite starter for food fermentation, is characterized in that: Karst Brettanomyces (Brettanomycescustersii) ZSM-001 of Rhizopus oryzae (Rhizopus oryzae) zsm-004 that this microbial composite starter includes Rhizopus oryzae (Rhizopus oryzae) zsm-003 that preserving number is CCTCC NO:M 2014374, preserving number is CCTCC NO:M2014375, preserving number to be head mold (Rhizopus sp.) zsm-005 of CCTCC NO:M 2014376 and preserving number be CCTCC NO:M207150.
5. the application of Rhizopus oryzae according to claim 1 (Rhizopus oryzae) zsm-003 in preparation microbial composite starter.
6. the application of Rhizopus oryzae according to claim 2 (Rhizopus oryzae) zsm-004 in preparation microbial composite starter.
7. the application of head mold according to claim 3 (Rhizopus sp.) zsm-005 in preparation microbial composite starter.
8. the application of complex microbial inoculum according to claim 4 in rice food.
9. apply as claimed in claim 8, comprising the application in sweet wine fermentation.
10. apply as claimed in claim 8, comprising the application in fermented rice cake fermentation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410676014.5A CN104357337B (en) | 2014-11-23 | 2014-11-23 | Rhizopus for food fermentation and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410676014.5A CN104357337B (en) | 2014-11-23 | 2014-11-23 | Rhizopus for food fermentation and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104357337A true CN104357337A (en) | 2015-02-18 |
| CN104357337B CN104357337B (en) | 2017-04-12 |
Family
ID=52524651
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410676014.5A Active CN104357337B (en) | 2014-11-23 | 2014-11-23 | Rhizopus for food fermentation and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104357337B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106754407A (en) * | 2016-12-02 | 2017-05-31 | 贵州省烟草科学研究院 | A kind of Rhizopus oryzae and application thereof |
| CN108485992A (en) * | 2018-06-06 | 2018-09-04 | 江西省食品发酵研究所 | A kind of head mold of food fermentation and application |
| CN113528350A (en) * | 2020-04-13 | 2021-10-22 | 安琪酵母股份有限公司 | Rhizopus strain, distiller's yeast and rice wine and production method of distiller's yeast |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN86103877A (en) * | 1986-06-03 | 1988-03-23 | 张雪岳 | Leaven of special flavour sweet wine |
| CN102041234A (en) * | 2009-10-22 | 2011-05-04 | 安琪酵母股份有限公司 | Rhizopus strains, yeast strains, distiller's yeast containing same and production method for distiller's yeast |
| CN102994396A (en) * | 2009-10-22 | 2013-03-27 | 安琪酵母股份有限公司 | Rhizopus oryzae strains, rice distiller's yeasts, production of rice distiller's yeasts, rice wine and production method of rice wine |
-
2014
- 2014-11-23 CN CN201410676014.5A patent/CN104357337B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN86103877A (en) * | 1986-06-03 | 1988-03-23 | 张雪岳 | Leaven of special flavour sweet wine |
| CN102041234A (en) * | 2009-10-22 | 2011-05-04 | 安琪酵母股份有限公司 | Rhizopus strains, yeast strains, distiller's yeast containing same and production method for distiller's yeast |
| CN102994396A (en) * | 2009-10-22 | 2013-03-27 | 安琪酵母股份有限公司 | Rhizopus oryzae strains, rice distiller's yeasts, production of rice distiller's yeasts, rice wine and production method of rice wine |
Non-Patent Citations (1)
| Title |
|---|
| 李健容等: "民间传统酒曲主要微生物的分离及鉴定", 《酿酒科技》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106754407A (en) * | 2016-12-02 | 2017-05-31 | 贵州省烟草科学研究院 | A kind of Rhizopus oryzae and application thereof |
| CN106754407B (en) * | 2016-12-02 | 2020-05-08 | 贵州省烟草科学研究院 | Rhizopus oryzae and application thereof |
| CN108485992A (en) * | 2018-06-06 | 2018-09-04 | 江西省食品发酵研究所 | A kind of head mold of food fermentation and application |
| CN113528350A (en) * | 2020-04-13 | 2021-10-22 | 安琪酵母股份有限公司 | Rhizopus strain, distiller's yeast and rice wine and production method of distiller's yeast |
| CN113528350B (en) * | 2020-04-13 | 2022-11-22 | 安琪酵母股份有限公司 | Rhizopus strain, distiller's yeast and rice wine and production method of distiller's yeast |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104357337B (en) | 2017-04-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107475012B (en) | Production method for brewing fen-flavor liquor by multi-strain enhanced Daqu fermentation | |
| CN104388514B (en) | The method that gamma aminobutyric acid is prepared using composite bacteria fermentation | |
| CN105349444B (en) | The saccharomycete of high yield ethyl acetate and its application under one plant of cryogenic conditions | |
| CN102140427B (en) | Method for preparing intensified daqu applied to nongjiang-flavor Chinese spirits | |
| CN111961615B (en) | Saccharopolyspora capable of reducing biogenic amine and application thereof | |
| CN102102084A (en) | Issatchenkia orientalis and composition and application thereof | |
| CN106701518A (en) | Block koji strengthening method capable of reducing dosage of block koji and improving quality of vinegar | |
| CN110205253A (en) | A kind of low yield isoamyl alcohol, the yeast of high yield bata-phenethyl alcohol and its isolated culture method and application | |
| CN106591160A (en) | Compound Xiaoqu and Xiaoqu Baijiu production method | |
| CN109971657B (en) | Rhizopus oryzae capable of producing saccharifying enzyme at high yield and application of rhizopus oryzae | |
| CN104357337B (en) | Rhizopus for food fermentation and application | |
| CN104430681B (en) | A kind of rice tailored flour for bread and manufacture method and application | |
| CN113249268B (en) | Saccharopolyspora rosea for reducing biogenic amine and application thereof | |
| CN105400656B (en) | A kind of brewing method of yellow rice wine | |
| CN102168027B (en) | New strain J4 for biofermentation of fruit wine and application thereof | |
| CN112011467B (en) | Rhizopus oryzae, microbial inoculum, bran koji, preparation methods thereof, application of rhizopus oryzae, microbial inoculum and bran koji, wine and preparation method of wine | |
| CN105002102A (en) | Kluyveromyces marxianus and cultural method thereof and application | |
| CN109566995B (en) | Pearl barley fermented glutinous rice can and preparation method thereof | |
| CN110218663B (en) | Bacillus amyloliquefaciens Q4 strain for high yield of alpha-glycosidase inhibitor, functional yellow wine and preparation method | |
| CN104250618B (en) | The aspergillus candidus of a kind of high-yield glucoamylase, alpha amylase and acid protease and its application | |
| CN105670936B (en) | A kind of method of Trametes trogii bacterial strain and its application and Pu'er tea processing | |
| CN109988721A (en) | One plant of methamidophos strain that can increase sauce based food fragrance | |
| CN113151005B (en) | Monascus purpureus W-4 capable of producing lovastatin at high yield and application thereof | |
| CN104762171B (en) | blueberry wine and preparation method thereof | |
| CN102604840B (en) | Rice wine yeast, rhizopus and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |