CN103060244B - Bacillus marinus and method for producing catalase by using same - Google Patents
Bacillus marinus and method for producing catalase by using same Download PDFInfo
- Publication number
- CN103060244B CN103060244B CN201310022577.8A CN201310022577A CN103060244B CN 103060244 B CN103060244 B CN 103060244B CN 201310022577 A CN201310022577 A CN 201310022577A CN 103060244 B CN103060244 B CN 103060244B
- Authority
- CN
- China
- Prior art keywords
- catalase
- cgmcc
- water
- strain
- hydrogen peroxide
- 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.)
- Active
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
The invention relates to an ocean source catalase high-yielding strain and a method for producing catalase by using the strain. One strain of Oceanobacillus iheyensis is obtained by directionally enriching and sieving from an ocean water body through adopting a semi-solid culture medium; and the preservation number is CGMCC (China General Microbiological Culture Collection Center) No. 6042. The strain is fermented to obtain the catalase and the enzyme activity reaches 8677U/mg. A preparation process of the catalase is simple; the enzymatic activity is high and the cost is low; and the catalase is suitable for being widely applied to the fields of foods, spinning, papermaking, environmental friendliness and the like. Therefore, an enzyme producing strain and a fermentation method disclosed by the invention have wide industrial application values and obvious economic benefit prospects.
Description
Technical field
The invention belongs to marine biotechnology technical field, be specifically related to a kind of marine source Catalase-Producing Strain and adopt this bacterium to produce catalatic method.
Background technology
Catalase is the terminal oxidase being extensively present in animal, plant and microbe, and it is with hydrogen peroxide (H
2o
2) be substrate, be decomposed into water and oxygen.Catalase can be removed rapidly the oxyradical producing in the redox reaction of bio-metabolic process, avoids cell injured by strong oxidizer, is important Antioxidative system in organism.In recent years, along with H
2o
2in the generally application of the industries such as weaving, papermaking, food, medicine and environmental protection, market is also extensively and rising tendency catalatic demand.
In papermaking and textile industry, catalase can be used for H
2o
2deoxidizing purification after bleaching, fine toxicity and the problem of environmental pollution that has solved traditional chlorophenols compound SYNTHETIC OPTICAL WHITNER of the method, can save water and the energy again; In foodstuffs industry, catalase can be used for using H in the decomposing food course of processing
2o
2residual as SYNTHETIC OPTICAL WHITNER, oxygenant, sanitas, also can be used for food oxygen scavenger and raising agent; Aspect medical, when can be used for producing and using medical apparatus, decompose H residual in sterilization and disinfection process
2o
2; Aspect environment protection, H
2o
2can be used for processing various trade effluents with catalase, be most commonly used to sulfur-bearing, containing the processing of phenol and cyanide wastewater.
At present, catalatic production is raw material mainly with the liver of the various animals such as pig, ox, sheep, through self-dissolving, process and to make wherein catalase stripping, this method be subject to that starting material restriction is serious, cost is higher, separation purifying technique is complicated, and have animal to take viruliferous hidden danger.Catalase extensively exists in supporting well microorganism, uses microorganism fermentative production catalase, compares with traditional processing technology, the advantage such as have that production of enzyme is large, technique is simple and production cost is low.External having produced catalatic commercially produced product with microbial fermentation processes at present, and the domestic related products that yet there are no.
The ecotope that ocean is special, has bred a large amount of microorganisms and enzyme resource thereof.And the bacterial strain majority of the fermentation using bacteria of authorizing at present product catalase patent is located away from pedotheque.Only Chinese patent 200810057969.7 provides the catalatic bacterial strain of high-yield of low-temperature of a kind of Antarctic Ocean water separation, and the catalase activity of this bacterial strain is 1200U/mg.
Summary of the invention
An object of the present invention is to provide a kind of energy and produce catalatic high yield bacterial strain.
Bacterial strain of the present invention is the catalase zymogenic bacteria that a strain adopts the screening of semisolid medium screening method to obtain, Classification And Nomenclature is Yi Ping room submarine ridge bacillus marinus (Oceanobacillus iheyensis), separated from ocean seawater sample, be preserved in Chinese common micro-organisms culture presevation administrative center, deposit number is CGMCC No.6042, preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica (100101), and preservation date is on April 24th, 2012.
Through identification of morphology and biological characteristic research, preserving number is that the bacterial strain of CGMCC No.6042 has following characteristics:
(1) colonial morphology: bacterium colony is rounded, smooth surface, edge is smooth, microprotrusion, oyster white;
(2) cellular form: gram-positive microorganism, under microcytoscope, (Olympus, BX40) presents mobility, and form is shaft-like;
(3) physiological and biochemical property: aerobic growth; Oxidase positive; Hydrolysis Vitamin C2, polysorbate40 or polysorbate60, be not hydrolyzed tween 80, DNA or urea; Do not produce indoles; Can utilize glucose, seminose, fructose or maltose to produce acid, can not utilize pectinose or rhamnosyl to produce acid; Responsive to amoxycilline Trihydrate bp, penbritin, paraxin, erythromycin, Vulkamycin. PA-93 or Benzylpenicillin, insensitive to nysfungin.
The 16S rRNA gene that is CGMCC No.6042 bacterial strain to preserving number carries out pcr amplification and sequencing, and recording 16S rRNA Gene Partial fragment length is 542bp(Seq ID No.1), the 16S rRNA sequence of CGMCC No.6042 bacterial strain is specifically shown in sequence table.The 16S rRNA gene order similarity of bacterial strain CGMCC No.6042 and known Yi Ping room submarine ridge bacillus marinus reference culture (O.iheyensis) is 100%.On source, the submarine ridge bacillus marinus reference culture separation of Yi Ping room is from eastern China sea east, and bacterial strain CGMCC No.6042 separation is from Middle Pacific seamount district; In physiological characteristic, the growth optimum pH of Yi Ping room submarine ridge bacillus marinus reference culture is 8.0-8.5, and the growth optimum pH of bacterial strain CGMCC No.6042 is 7.0-7.5.Therefore, bacterial strain CGMCC No.6042 and Yi Ping room submarine ridge bacillus marinus reference culture are different strains of the same race.
Therefore, content with reference to < < Bergey ' s Manual of Systematic Bacteriology > > second edition, according to bacterial strain limiting factor feature, morphological specificity and physiological and biochemical index, in conjunction with 16S rRNA gene order similarity, identify that bacterial strain CGMCC No.6042 is Yi Ping room submarine ridge bacillus marinus (O.iheyensis).
The CGMCC No.6042 bacterial strain that another object of the present invention is to provide described in a kind of utilization is produced catalatic method.
Adopt O.iheyensis CGMCC No.6042 to produce catalase, with CGMCC No.6042 bacterial strain, for the bacterial classification that sets out, through seed culture and liquid fermenting, obtain catalase, comprise the following steps:
(1) bacterial strain CGMCC No.6042 is seeded to natural sea-water liquid nutrient medium, after cultivation, adds hydrogen peroxide to stimulate and cultivate as seed liquor;
(2) seed liquor is forwarded in fermention medium and carries out fermentation culture, after cultivation, add hydrogen peroxide to stimulate and cultivate, obtain having the fermented liquid of catalase activity.
In aforesaid method step (1), the formula of described natural sea-water substratum is in natural sea-water, to add the casamino acids of the peptone of 0.1-0.5%, the yeast extract of 0.01-0.05% and 0.1-0.5%, and adjusting pH is 6.8-7.5.In a preferred embodiment, natural sea-water liquid nutrient medium is in natural sea-water, to add 0.3% peptone, 0.02% yeast extract, and 0.3% casamino acids, regulating pH is 7.2.
In step (1), the microbial culture time is 12-48 hour, preferably 30-40 hour; Culture temperature is 20-40 ℃, preferably 30-35 ℃.After cultivation finishes, add the hydrogen peroxide of 1-10mM to stimulate cultivation 1-10 hour, preferably add the hydrogen peroxide of 2-5mM to stimulate cultivation 2-5 hour.
In aforesaid method step (2), described fermentative medium formula is in natural sea-water, to add the yeast extract of 0.1-1% and the peptone of 0.5-5%, and pH is 6.5-7.5.In a preferred embodiment, fermentative medium formula is in natural sea-water, to add the yeast extract of 0.4-0.6% and the peptone of 0.1-0.2%, and pH is 6.5-7.5.Incubation time is 12-48 hour, preferably 40-48 hour; Culture temperature is 20-40 ℃, preferably 30-35 ℃.After cultivation finishes, add the hydrogen peroxide of 1-10mM to stimulate cultivation 1-10 hour, preferably add the hydrogen peroxide of 2-5mM to stimulate cultivation 2-5 hour.
According to aforesaid method, can obtain the fermented liquid that activity of catalase is 1000-10000U/mg, enzyme activity reaches 8677U/mg in a preferred embodiment.
As required, can optionally to having the fermented liquid of enzyme activity, process, therefrom separation and purification goes out catalatic product, and preferred purification condition is as ion exchange chromatography, reversed phase chromatography, hydrophobic chromatography, sieve chromatography or its combination.
The O.iheyensis CGMCC No.6042 that the present invention finds can express highly active catalase, through hydrogen peroxide, stimulates, and expression amount and the activity of enzyme significantly improve.Catalase preparation technology provided by the invention is simple, and enzymic activity is high, with low cost, is applicable to being widely used in the fields such as food, weaving, papermaking and environmental protection.Therefore bacterium producing multi enzyme preparation of the present invention and fermentation process have industrial application value and significant economic benefit prospect widely.
Embodiment
The separation screening of embodiment 1 Yi Ping room submarine ridge bacillus marinus (O.iheyensis) CGMCC No.6042
4.5mL natural sea-water is adopted to NaOH or HCl fine setting pH to 7.2, add 0.75%(w/v) agar and in 121 ℃ of high pressure moist heat sterilizations 20 minutes, treat that temperature is reduced to 50 ℃ of left and right, the 30% hydrogen peroxide 4 μ L that add filtration sterilization under aseptic condition, mix rear cooled and solidified and make seawater semisolid medium.Under aseptic condition, the seawater sample that adds 4.5mL to gather in seawater semisolid medium, and the carbon and nitrogen sources of filtration sterilization, final concentration (w/v) is peptone 0.25%, yeast extract 0.01% and casamino acids 0.25%.28 ℃ of concussion enrichment culture, about 4 days, are diluted suitable multiple spread plate by pregnant solution, and picking mono-clonal carries out inclined-plane preservation.
The bacterial strain that aforesaid method separation obtains is forwarded to natural sea-water liquid nutrient medium and carries out shake flask fermentation, adopts spectrophotometry to carry out catalase activity and sieves again.Natural sea-water substratum is in natural sea-water, to add 0.25% peptone, 0.01% yeast extract and 0.25% casamino acids, and regulating pH is 7.2.
Spectrophotometry enzymic activity is linear with its concentration based on hydrogen peroxide light absorption value size under 240nm.Measure with method of calculation as follows: (1) is broken born of the same parents' processing by bacterium liquid and obtained crude enzyme liquid; (2) enzyme reaction system is 300 μ L, comprises crude enzyme liquid 5 μ L, phosphoric acid buffer 195 μ L and 10mM H
2o
2solution 100 μ L, substitute H with deionized water
2o
2solution is as blank; With H
2o
2solution add startup enzymatic reaction; Under (3) 30 ℃ of conditions, every 5s, read absorbance one time, minute is 1min, usings the rate of descent of absorbancy under 240nm wavelength as calculating the enzyme big or small foundation of living.Standard enzyme unit (1U) that lives is defined as: at 30 ℃, and the per minute 1 μ mol H that degrades
2o
2(1 μ mol/min) required enzyme amount.Enzyme work is calculated: (Δ OD
240/ Δ t) * 1000 * extension rate * reaction system volume/(43.6 * enzyme liquid is long-pending), 43.6M
-1cm
-1for H
2o
2molar extinction coefficient under 240nm.
By aforesaid method, screened and obtained the relatively high bacterial strain of a strain activity of catalase, be preserved in Chinese common micro-organisms culture presevation administrative center, culture presevation number is CGMCC No.6042.
Identification of morphology and the biological characteristics of embodiment 2 Yi Ping room submarine ridge bacillus marinus (O.iheyensis) CGMCC No.6042
Yi Ping room submarine ridge bacillus marinus (O.iheyensis) CGMCC No.6042 is inoculated in natural sea-water liquid nutrient medium, prepares the agar that solid medium can add 20g/L again.After cultivation, through identifying, this bacterium has following characteristics: (1) colonial morphology: bacterium colony is rounded, and smooth surface, edge is smooth, microprotrusion, oyster white; (2) cellular form: gram-positive microorganism, under microcytoscope, (Olympus, BX40) presents mobility, and form is shaft-like; (3) physiological and biochemical property: aerobic growth; Oxidase positive; Hydrolysis Vitamin C2, polysorbate40 or polysorbate60, be not hydrolyzed tween 80, DNA or urea; Do not produce indoles; Can utilize glucose, seminose, fructose or maltose to produce acid, can not utilize pectinose or rhamnosyl to produce acid; Responsive to amoxycilline Trihydrate bp, penbritin, paraxin, erythromycin, Vulkamycin. PA-93 or Benzylpenicillin, insensitive to nysfungin.
Pcr amplification and the sequencing of the 16S rRNA gene of embodiment 3 Yi Ping room submarine ridge bacillus marinus (O.iheyensis) CGMCC No.6042
Yi Ping room submarine ridge bacillus marinus (O.iheyensis) CGMCC No.6042 is inoculated in natural sea-water solid medium, from inclined-plane, directly picking one encircles thalline, add 200 μ L sterilized waters abundant suspension thalline, 100 ℃ of water-baths 30 minutes, centrifugal 2 minutes of 12000rpm immediately ice bath, supernatant liquor carries out PCR as template.The a pair of universal primer of amplification 16S rRNA gene is as follows:
Forward primer: 5'-AGAGTTTGATCCTGGCTCAG-3';
Reverse primer: 5'-GGTTACCTTGTTACGACTT-3 ';
Above-mentioned primer is 8-27 and the 1510-1492 bit base of corresponding colibacillary 16S rRNA gene respectively.PCR reaction system (50 μ L) is: 10 * buffer, 5 μ L, 10mM dNTPs1 μ L, each 1 μ L of 4 μ M primers, Taq enzyme 0.5 μ L, template DNA 0.5 μ L, sterile pure water 41 μ L.PCR reaction conditions is: 94 ℃ of denaturation 5min; 94 ℃ of sex change 45s, 55 ℃ of annealing 45s; 72 ℃ are extended 90s, 30 circulations; After 72 ℃, extend 10min.PCR product purification and sequencing are completed by Shanghai Sheng Gong biotechnology company limited.The 16S rRNA Gene Partial fragment length that completes mensuration is 542bp, with bacterial strain O.iheyensis JCM11309
t16S rRNA gene order similarity the highest.The concrete visible sequence table of 16S rRNA sequence of bacterial strain O.Iheyensis CGMCC No.6042.
Therefore, with reference to < < Bergey ' s Manual ofSystematic Bacteriology > > second edition content, according to limiting factor feature, morphological specificity and the physiological and biochemical index of bacterial strain, in conjunction with 16S rRNA gene order similarity, identify that CGMCC No.6042 is Yi Ping room submarine ridge bacillus marinus (O.iheyensis).
Embodiment 4 fermention mediums and fermentation time optimization
Yi Ping room submarine ridge bacillus marinus (O.iheyensis) CGMCC No.6042 is seeded to natural sea-water liquid nutrient medium, cultivates 36h for 28 ℃ and reaches logarithmic growth latter stage.Add the hydrogen peroxide of 4mM to stimulate cultivation 2h as seed liquor.Be forwarded to the 250mL triangular flask that 50mL fermention medium is housed and carry out fermentation culture.Fermented incubation time is 24,36 and 48h, 28 ℃ of culture temperature, shaking speed 200r/min.Fermention medium has three kinds: (1) seawater fermention medium: 0.1% peptone, and 0.5% yeast extract, all the other are natural old seawater, pH7.2; (2) extractum carnis fermention medium: 0.5% extractum carnis, 1% peptone, 0.5% yeast extract, 2.3% sodium-chlor, pH7.2; (3) casein food grade-glucose fermentation substratum: 2% casein food grade, 4% glucose, 0.1% potassium primary phosphate, 0.01% magnesium sulfate, 2.3% sodium-chlor, pH7.2.By three kinds of different fermentations substratum and time, cultivated and collected thalline, after broken born of the same parents, measure catalase enzyme activity, concrete outcome is in Table 1.
The catalase activity that table 1 fermention medium and fermentation time optimization are measured afterwards
Yi Ping room submarine ridge bacillus marinus (O.iheyensis) CGMCC No.6042 is through seawater fermention medium fermentation culture 24-48h, and its activity of catalase value is first high rear low, is then increased to 3289.0U/mL again.Through extractum carnis substratum fermentation culture 24-48h, it is low that the variation of O.iheyensis CGMCC No.6042 activity of catalase value presents two ends, middle high feature; Enzyme activity maximum value appears at fermentation culture 36h, is 1087.2U/mL.Through casein food grade-dextrose culture-medium fermentation culture 24-48h, O.iheyensis CGMCC No.6042 activity of catalase value reaches maximum value (1678.9U/mL) when yeast culture 24h, then with the yeast culture time, increases enzyme decline gradually alive.Consider fermentation costs and production efficiency, Yi Ping room submarine ridge bacillus marinus (O.iheyensis) CGMCC No.6042 catalase fermentative production preferentially selects seawater fermention medium to cultivate 48h, secondly be casein food grade-glucose fermentation culture medium culturing 24h, again for extractum carnis fermention medium is cultivated 36h.
In embodiment 5 fermenting processs, adopting hydrogen peroxide to stimulate increases activity of catalase
Yi Ping room submarine ridge bacillus marinus (O.iheyensis) CGMCC No.6042 is seeded to natural sea-water liquid nutrient medium, culture condition is 28 ℃ and cultivates 36h as seed liquor, transfer respectively contain 0,0.25,0.5,1.0 and the natural sea-water liquid nutrient medium of 2.0mM hydrogen peroxide in cultivate, determine that the highest concentration of hydrogen peroxide that bacterial strain can be grown is 1.0mM.
The O.Iheyensis CGMCC No.6042 bacterium liquid of growing under 1mM concentration of hydrogen peroxide is forwarded to the 250mL triangular flask that 50mL seawater fermention medium is housed and carries out fermentation culture, incubation time 48h, 28 ℃ of culture temperature, shaking speed 200r/min.Cultivation finishes the rear hydrogen peroxide that adds respectively sterile pure water and 4mM to stimulate and cultivates the mensuration of carrying out catalase activity after 1h.
The fermentation broth enzyme stimulating without hydrogen peroxide is lived as 3269U/mL(5133U/mg), through hydrogen peroxide, stimulate the fermentation broth enzyme work obtaining to reach 5527U/mL(8677U/mg).Utilize hydrogen peroxide to stimulate to improve the method effect of bacterial strain catalase activity more remarkable, enzyme activity increase rate reaches 69%(3544U/mg).
Claims (6)
1. a catalase zymogenic bacteria, Classification And Nomenclature is Yi Ping room submarine ridge bacillus marinus (Oceanobacillus iheyensis), deposit number is CGMCC No.6042.
2. utilize the bacterial strain described in claim 1 to produce a catalatic method, comprise the following steps:
(1) bacterial strain CGMCC No.6042 is seeded to natural sea-water liquid nutrient medium, after cultivation, adds hydrogen peroxide to stimulate and cultivate as seed liquor; The formula of described natural sea-water substratum is in natural sea-water, to add the casamino acids of the peptone of 0.1-0.5%, the yeast extract of 0.01-0.05% and 0.1-0.5%, and adjusting pH is 6.8-7.5;
(2) seed liquor is forwarded in fermention medium and carries out fermentation culture, after cultivation, add hydrogen peroxide to stimulate and cultivate, obtain having the fermented liquid of catalase activity; Described fermentative medium formula is in natural sea-water, to add the yeast extract of 0.1-1% and the peptone of 0.5-5%, and pH is 6.5-7.5.
3. method according to claim 2, is characterized in that: in described method steps (2), microbial culture adds the hydrogen peroxide of 1-10mM to stimulate cultivation 1-10 hour after finishing.
4. method according to claim 2, is characterized in that: the activity of catalase that described method steps (2) obtains in fermented liquid is 1000-10000U/mg.
5. according to the method described in claim 2-4 any one, it is characterized in that: to having the fermented liquid of enzyme activity, process, therefrom separation and purification goes out catalase product.
6. method according to claim 5, is characterized in that: described separation and purification is ion exchange chromatography, reversed phase chromatography, hydrophobic chromatography, sieve chromatography or its combination.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310022577.8A CN103060244B (en) | 2013-01-21 | 2013-01-21 | Bacillus marinus and method for producing catalase by using same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310022577.8A CN103060244B (en) | 2013-01-21 | 2013-01-21 | Bacillus marinus and method for producing catalase by using same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103060244A CN103060244A (en) | 2013-04-24 |
| CN103060244B true CN103060244B (en) | 2014-04-23 |
Family
ID=48103144
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310022577.8A Active CN103060244B (en) | 2013-01-21 | 2013-01-21 | Bacillus marinus and method for producing catalase by using same |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103060244B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103789225A (en) * | 2013-11-08 | 2014-05-14 | 浙江大学 | Marine catalase production strain and method for producing catalase from strain |
| CN103952347B (en) * | 2014-04-22 | 2016-01-06 | 四川大学 | One strain moderate salt tolerant bacillus marinus and application thereof |
| CN103966146B (en) * | 2014-05-28 | 2016-03-30 | 四川大学 | One strain reaches wooden bacillus marinus and application thereof |
| CN111440807A (en) * | 2020-04-07 | 2020-07-24 | 上海海洋大学 | Shewanella WP3 mutant strain with high yield of low-temperature catalase as well as construction method and application thereof |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101230338B (en) * | 2008-02-22 | 2010-08-25 | 中国水产科学研究院黄海水产研究所 | Isolation and purification method for high-yield of low-temperature catalase by antarctic marine bacillus n2a |
| CN101245331B (en) * | 2008-02-22 | 2011-06-22 | 中国水产科学研究院黄海水产研究所 | South pole marine microorganism bacterial strain n2a of high-production low-temperature catalase |
| CN102864106B (en) * | 2012-09-24 | 2014-04-23 | 国家海洋局第二海洋研究所 | Ocean catalase production bacteria and directional screening method |
-
2013
- 2013-01-21 CN CN201310022577.8A patent/CN103060244B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN103060244A (en) | 2013-04-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102703339B (en) | High-yield arginine deiminase bacterial strain and method for producing L-citrulline by same | |
| CN109153966A (en) | Produce the biological technique method and related new strains of acrylamide | |
| CN103060244B (en) | Bacillus marinus and method for producing catalase by using same | |
| JP2012239452A (en) | Highly starch-accumulating microalgae, method for producing glucose using the same, and method for producing objective substance | |
| CN102864113B (en) | Strain capable of producing succinic acid, method for producing succinic acid and application thereof | |
| CN104630166A (en) | Method for producing low-temperature glucose oxidase by virtue of microbial fermentation | |
| CN103789225A (en) | Marine catalase production strain and method for producing catalase from strain | |
| CN104630167A (en) | Method for producing low-temperature glucose oxidase by fermentation of marine microorganisms | |
| CN102864106B (en) | Ocean catalase production bacteria and directional screening method | |
| CN110129225A (en) | γ~polyglutamic acid producing strains and breeding prepare γ~polyglutamic acid method | |
| CN105861373B (en) | It is a kind of produce keratinase pseudomonas aeruginosa and its application | |
| TWI598442B (en) | Medium For Producing Glucosamine And Its Application | |
| CN101153297A (en) | Novel single-tank hemicontinuous high-strength ferment high optical purity L- lactic acid technique for rhizopus oryzae bacterium ball | |
| JP2020054334A (en) | Heterotrophic bacteria cupriavidus gilardii gbs-15-1 strain, which is associate for obtaining microbial protein | |
| CN114015607B (en) | Bacillus amyloliquefaciens for high yield of 5-methyltetrahydrofolic acid and application thereof | |
| CN109161507A (en) | A kind of Corynebacterium glutamicum of high yield L-Orn and its application | |
| CN101463370B (en) | Method for preparing L-lactic acid by fermenting potato starch by Rhizopus oryzae | |
| CN103667107A (en) | Enterococcus faecium strain capable of producing L-lactic acid | |
| CN109161570B (en) | Method for improving fermentation production of N-acetylneuraminic acid and fermentation liquor | |
| CN112359074A (en) | Method for stimulating heterotrophic microalgae oil production by using acetic acid | |
| CN101659970A (en) | Method for circularly treating avermectins waste ferment water and pleurin waste ferment water | |
| CN101497871A (en) | Alcohol fermentation anaerobic high temperature bacterium culture medium, preparation and use thereof | |
| CN105368911A (en) | Method for synthesizing polyhydroxyalkanoate with kitchen waste anaerobic fermentation liquid as substrate | |
| RU2175014C2 (en) | Method of lactic acid producing | |
| KR101173468B1 (en) | Enterobacter sp?ES392 KACC 91568P and method of producing hydrogen by using the same |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |