CN103146661A - Method for improving yield of glutamine transaminase of streptoverticillium mobaraense in forced manner by MgCl2 - Google Patents
Method for improving yield of glutamine transaminase of streptoverticillium mobaraense in forced manner by MgCl2 Download PDFInfo
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- CN103146661A CN103146661A CN2013100720243A CN201310072024A CN103146661A CN 103146661 A CN103146661 A CN 103146661A CN 2013100720243 A CN2013100720243 A CN 2013100720243A CN 201310072024 A CN201310072024 A CN 201310072024A CN 103146661 A CN103146661 A CN 103146661A
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Abstract
The invention discloses a method for improving the yield of glutamine transaminase of streptoverticillium mobaraense in a forced manner by MgCl2, and relates to the technical field of production of glutamine transaminase by fermentation of streptomycete. According the method provided by the invention, streptoverticillium mobaraense is used as a fermenting bacterial strain which is inoculated to a fermenting culture medium after bevel culture and seed culture so as to prepare glutamine transaminase by liquid fermentation. The target of improving the yield of glutamine transaminase is finally realized by adding MgCl2 into the fermenting culture medium, and a protective preparation is added to prepare a low temperature spray drying or freeze drying powder. MgCl2 used in the invention is low in cost, and the yield of glutamine transaminase is remarkably improved. Under the condition that other culture conditions are same, the yield is improved by about 100% compared with that of a reference product without MgCl12, and the fermenting period is shortened.
Description
Technical field
The present invention relates to streptomycete fermentation and produce the glutamine transaminage technical field, be specifically related to a kind of interpolation MgCl
2Improve the method that fermentation method prepares glutamine transaminage.
Background technology
Glutamine transaminage (Transglutaminase; be called for short TGase; EC 2.3.2.13; R-glutaminyl-peptide:amnie-γ-glutamyl-transferase) has another name called trans-glutaminases; it carries out the acyl group shift reaction by the methane amide on the glutaminic acid residue in catalytic proteins and the epsilon-amino on lysine residue, make protein in molecule or intermolecular formation crosslinked.
In the 1980s, the people such as Ikura have reported the crosslinked action of glutamine transaminage to food proteins.They are with the crosslinked several food proteinss of mammalian source glutamine transaminage, as casein, soybean protein etc.These albumen crosslinked improved the functional of its product, as gelation, solvability and emulsifying property etc.Yet, because the utilization ratio of its fancy price and poor efficiency has limited its application in foodstuffs industry.Successfully being separated to the microbial source glutamine transaminage from streptomycete is to promote its milestone using in foodstuffs industry.It makes this kind of enzyme to produce on a large scale, and by commercial exploitation.Catalysis characteristics due to streptomycete glutamine transaminage substrate, and its broad prospect of application in food, weaving and pharmaceutical industries, be subject to the extensive concern of Chinese scholars, Japan, Taiwan in the Asia, the Denmark, Holland, France, Germany, Ireland in Europe and the country such as Brazilian have all dropped into a large amount of human and material resources, financial resources are studied, and have obtained the progress of advancing by leaps and bounds.
The source of glutamine transaminage mainly contains 3 kinds: the one, extract from the tissue of animal or body fluid, such as ox, pig or fish etc., limited due to animal tissues's kind, the source is rare, content is low, tissue specificity is high again, and the separation and purification process is complicated, price is very expensive; In Europe; business-like factor XIII (glutamine transaminage a kind of) mainly obtains by slaughtering the animals such as ox or pig; but because needing thrombin activation usually, the glutamine transaminage in blood just has activity, therefore seldom be applied in foodstuffs industry.The 2nd, take intestinal bacteria, coryneform bacteria and streptomycete etc. as the host, produce this enzyme with genetic engineering means.The genetic engineering bacterium expression effect is relatively poor, far can not reach previous level; Although some report output is higher, some country is stricter to the gene engineering product restriction ratio.The 3rd, directly the microorganism of glutamine transaminage is produced in screening, produces by fermentation method.Lower due to this method technical feasibility, cost, produce do not set out material and season limit, most possibly carry out scale operation and widespread use.But at present food grade glutamine transaminage source is single, output is lower, expensive, has limited its application in foodstuffs industry.
The report that improves glutamine of microbe transaminase output both at home and abroad concentrates on optimization of fermentation conditions mostly, improves the aspects such as substratum composition.Although these researchs have all obtained some achievements, the output of this enzyme is still lower.Present business-like product is mainly produced by companies such as Japanese aginomotos.Domesticly still be in the starting stage with the Production by Microorganism Fermentation glutamine transaminage, output is lower.
Summary of the invention
Problem for the preparation method who has glutamine transaminage now exists the invention provides a kind of MgCl
2Coerce the method that improves streptomyces mobaraensis glutamine transaminage output.
Goal of the invention of the present invention is achieved by the following technical solution:
The employing streptomyces mobaraensis is fermentation strain, after slant culture and seed culture, is seeded in fermention medium and prepares glutamine transaminage through liquid fermenting, by add MgCl in fermention medium
2, fermentation 96 h have finally realized improving the target of glutamine transaminage output.
Concrete technical scheme is as follows:
1, bacterial classification and substratum:
(1) bacterial classification:
Streptomyces mobaraensis (
Streptomyces mobaraense) DSM40587: be purchased from Japanese NBRC.
(2) slant medium (g/L): Gause I substratum.
(3) seed culture medium (g/L): polyprotein peptone 20, Zulkovsky starch 20, dipotassium hydrogen phosphate 2, potassium primary phosphate 2, yeast powder 2, anhydrous magnesium sulfate 2, pH 7.0.
(4) initial fermention medium (g/L): polyprotein peptone 30, Zulkovsky starch 10, fructose 10, dipotassium hydrogen phosphate 2, yeast powder 2, anhydrous magnesium sulfate 1, pH 7.0.
2, glutamine transaminage enzyme measuring method alive:
Adopt under 37 ℃ of spectrophotometrys and measure.Draw 50 μ L fermented liquids, add the 0.5mL reagent A, 37 ℃ of water bath heat preservation 10min add 0.5mL reagent B, termination reaction, 500nm colorimetric.
The N-CBZ-Gln-Gly of reagent A: 100mg is dissolved in the NaOH of 2mL, 0.2mol/L, adds successively the Tris-HCl damping fluid 4mL of 0.2mol/L, pH 6.0, the azanol 2mL of 0.1mol/L, and the reduced glutathion 2mL of 0.01mol/L, and regulate pH to 6.0.
Reagent B:3mol/L hydrochloric acid, 12% trichoroacetic acid(TCA), 5%FeCl
36H
2O(is dissolved in the hydrochloric acid of 0.1mol/L) mix in the ratio of 1:1:1.
1 unit glutamine transaminage active (U) is defined as: 37 ℃, react under the condition of pH 6.0, and it is the required enzyme amount of single hydroximic acid of 1 μ mol that per minute produces concentration.
3, cultural method:
The slant pore that to cultivate 7 d with the 5mL stroke-physiological saline solution washes, be inoculated in 100 mL seed culture mediums, after 30 ℃, 200 r/min are cultivated 48 h, add in fermention medium by 10% inoculum size, the training method of taking under 30 ℃, 200 r/min conditions to improve glutamine transaminage output is cultivated.Cultured fermented liquid is filtered or press filtration through plate filter; microfilter device through 0.22 μ m filter membrane filters again; make acellular enzyme liquid; add carrier protecting agent fully miscible, or these protective materials and acellular enzyme liquid fully mix the microfilter device filtration sterilization through 0.22 μ m filter membrane again.Adopt low temperature spray drying or lyophilize to make dry powder.Or by making the pharmaceutical grade zymin through freeze-drying again after ultrafiltration, gel chromatography, ion exchange chromatography, dialysis.
4, the training measures of above-mentioned raising glutamine transaminage output:
The training method of related raising glutamine transaminage output is to add 0.1-0.2 mol/LMgCl in initial fermention medium
2, in fermentation 96 h fermented liquids, the glutamine transaminage vigor reaches 4.3 U/mL.And not for adding MgCl
2Enzyme activity reaches maximum (2.1U/mL) when fermentation 120 h.MgCl
2Not only promoted the output of enzyme, and fermentation period shortens 24h.
5, pharmaceutical grade enzyme preparation method:
(1) ultrafiltration
Above-mentioned acellular enzyme liquid is carried out ultrafiltration, and the ultrafiltration condition is the ultra-filtration membrane of 30kDa for adopting the molecular weight that dams, with 1/5 of acellular enzyme liquid simmer down to original volume.
(2) gel chromatography
Concentrated solution is joined in the Sephadex G75 post of using in advance 50 mmol/L phosphate buffered saline buffers (pH 6.0) balance, and carry out wash-out with same buffer, flow velocity is 0.25 mL/min, detects wavelength for 280nm, collects activated part;
(3) ion exchange chromatography:
Whole active parts of collecting are joined in SP sepharose High Performance (HP) (1.6 * 10 cm) chromatography column of using in advance 50 mmol/L, pH 6.0 phosphate buffered saline buffer balances, carry out linear gradient elution with the same buffer that contains 0-0.25mol/L sodium-chlor, flow velocity is 3 mL/min, detect wavelength for 280nm, collect active part.
6, carrier protecting agent adds:
Above-mentioned related protective material is in acellular enzyme liquid sucrose 1-2%, trehalose 3-5%, glycerine 3-6%.Carrier protecting agent was through oven dry in 100 ℃, 1 hour, and under aseptic condition, mixer is fully miscible; Or these protective materials and acellular enzyme liquid are fully miscible again through the microfilter device filtration sterilization of 0.22 μ m filter membrane.
7, dry powder is made in low temperature spray drying or lyophilize:
After carrier is added protective material, adopt spray drying method in low temperature dry, condition is 80 ℃ of inlet temperature, air outlet temperature 55-60 ℃.Perhaps adopt vacuum freeze drier dry, the cold well temperature of Freeze Drying Equipment is-60 ℃, and vacuum tightness is 0.05MPa.
The present invention adopts MgCl
2Coerce and promote streptomyces mobaraensis to produce glutamine transaminage, the purified rear thermostability of the enzyme that this law obtains and pH stability are all higher than the synthetic glutamine transaminage of streptomyces hygroscopicus, and it has very large potentiality in food-processing, textile industry and organization bracket prepare industry.MgCl used
2Cheap, simple to operate, easy to implement, significantly improved the output of glutamine transaminage, in the situation that other culture condition is identical, the present invention is than not adding MgCl
2Reference examples output improve approximately 100%, and shortened fermentation period, reach the advanced level that fermentation method prepares glutamine transaminage.
Embodiment
Embodiment one: when fermentation is initial, add MgCl in culturing bottle
2Making its final concentration is 0.20 mol/L.Cultivate 96 h under 30 ℃, 200 r/min conditions, in fermented liquid, enzyme activity rises to 3.8U/mL by 0.6U/mL.Cultured fermented liquid is filtered or press filtration through plate filter, then filter through the microfilter device of 0.22 μ m filter membrane, make acellular enzyme liquid.Carrier protecting agent sucrose 1-2%, trehalose 3-5%, glycerine 3-6% was through oven dry in 100 ℃, 1 hour, and is fully miscible with aseptic enzyme liquid under aseptic condition; Or acellular enzyme liquid and carrier protecting agent sucrose 1-2%; trehalose 3-5%; glycerine 3-6% is fully miscible; through the microfilter device filtration sterilization of 0.22 μ m filter membrane, adopt spray drying method in low temperature dry again, condition is 80 ℃ of inlet temperature; air outlet temperature 55-60 ℃; perhaps adopt vacuum freeze drier dry, the cold well temperature of Freeze Drying Equipment is-60 ℃, and vacuum tightness is 0.05MPa.
Embodiment two: add 0.15 mol/LMgCl in initial fermented liquid
2, the enzyme work of fermentation 96h reaches 4.2U/mL, cultured fermented liquid is filtered or press filtration through plate filter, then filter through the microfilter device of 0.22 μ m filter membrane, makes acellular enzyme liquid.Carrier protecting agent sucrose 1-2%, trehalose 3-5%, glycerine 3-6% was through oven dry in 100 ℃, 1 hour, and is fully miscible with aseptic enzyme liquid under aseptic condition; Or acellular enzyme liquid and carrier protecting agent sucrose 1-2%; trehalose 3-5%; glycerine 3-6% is fully miscible; through the microfilter device filtration sterilization of 0.22 μ m filter membrane, adopt spray drying method in low temperature dry again, condition is 80 ℃ of inlet temperature; air outlet temperature 55-60 ℃; perhaps adopt vacuum freeze drier dry, the cold well temperature of Freeze Drying Equipment is-60 ℃, and vacuum tightness is 0.05MPa.
Embodiment three: add 0.1 mol/L MgCl in initial fermented liquid
2, fermentation 120h enzyme activity reaches maximum (4.3U/mL), cultured fermented liquid is filtered or press filtration through plate filter, then filter through the microfilter device of 0.22 μ m filter membrane, makes acellular enzyme liquid.Carrier protecting agent sucrose 1-2%, trehalose 3-5%, glycerine 3-6% was through oven dry in 100 ℃, 1 hour, and is fully miscible with aseptic enzyme liquid under aseptic condition; Or acellular enzyme liquid and carrier protecting agent sucrose 1-2%; trehalose 3-5%; glycerine 3-6% is fully miscible; through the microfilter device filtration sterilization of 0.22 μ m filter membrane, adopt spray drying method in low temperature dry again, condition is 80 ℃ of inlet temperature; air outlet temperature 55-60 ℃; perhaps adopt vacuum freeze drier dry, the cold well temperature of Freeze Drying Equipment is-60 ℃, and vacuum tightness is 0.05MPa.
Specific implementation method four: present embodiment and embodiment one, two, three different be that by making the pharmaceutical grade zymin through freeze-drying again after ultrafiltration, gel chromatography, ion exchange chromatography, dialysis, concrete steps are as follows:
(1) ultrafiltration
Above-mentioned acellular enzyme liquid is carried out ultrafiltration, and the ultrafiltration condition is the ultra-filtration membrane of 30kDa for adopting the molecular weight that dams, with 1/5 of acellular enzyme liquid simmer down to original volume.
(2) gel chromatography
Concentrated solution is joined in the Sephadex G75 post of using in advance 50 mmol/L phosphate buffered saline buffers (pH 6.0) balance, and carry out wash-out with same buffer, flow velocity is 0.25 mL/min, detects wavelength for 280nm, collects activated part;
(3) ion exchange chromatography
Whole active parts of collecting are joined in SP sepharose High Performance (HP) (1.6 * 10 cm) chromatography column of using in advance 50 mmol/L phosphate buffered saline buffers (pH 6.0) balance, carry out linear gradient elution with the same buffer that contains 0-0.25mol/L sodium-chlor, flow velocity is 3 mL/min, detect wavelength for 280nm, collect active part.Be the pure Transglutaminase EC2.3.2.13 of prepared electrophoresis.What the rate of recovery of TGASE can reach 77.5%, TGASE can reach 17.33U/mg than enzyme work.
Claims (10)
1. MgCl
2Coerce the method that improves streptomyces mobaraensis glutamine transaminage output, the employing streptomyces mobaraensis is fermentation strain, after slant culture and seed culture, be seeded in fermention medium and prepare glutamine transaminage through liquid fermenting, it is characterized in that: add MgCl in fermention medium
2 ,96 h that ferment improve the output of streptomycete glutamine transaminage.
2. MgCl according to claim 1
2Coerce the method that improves streptomyces mobaraensis glutamine transaminage output, it is characterized in that adding in described fermention medium the MgCl of 0.1-0.2 mol/L
2
3. MgCl according to claim 1
2Coerce the method that improves streptomyces mobaraensis glutamine transaminage output, it is characterized in that described MgCl
2Coerce the concrete steps that improve streptomyces mobaraensis glutamine transaminage output as follows:
The slant pore that to cultivate 7d with the 5mL stroke-physiological saline solution washes, be inoculated in the 100mL seed culture medium, 30 ℃, 200r/min add in fermention medium by 10% inoculum size after cultivating 48h, add 0.1-0.2 mol/L MgCl in initial fermention medium
2The training method of taking under 30 ℃, 200r/min condition to improve glutamine transaminage output is cultivated; cultured fermented liquid is filtered or press filtration through plate filter; microfilter device through 0.22 μ m filter membrane filters again; make acellular enzyme liquid; the carrier protecting agent that adds the drying sterilization is fully miscible, makes dry powder or pharmaceutical grade zymin.
4. MgCl according to claim 3
2Coerce the method that improves streptomyces mobaraensis glutamine transaminage output, it is characterized in that described carrier protecting agent is in acellular enzyme liquid sucrose 1-2%, trehalose 3-5%, glycerine 3-6%.
5. according to claim 3 or 4 described MgCl
2Coerce the method that improves streptomyces mobaraensis glutamine transaminage output, it is characterized in that described carrier protecting agent through 100 ℃, 1 hour drying and sterilizing, fully miscible with acellular enzyme liquid under aseptic condition.
6. according to claim 3 or 4 described MgCl
2Coerce the method that improves streptomyces mobaraensis glutamine transaminage output, it is characterized in that described protective material and acellular enzyme liquid are fully miscible, then through the microfilter device filtration sterilization of 0.22 μ m filter membrane.
7. MgCl according to claim 3
2Coerce the method that improves streptomyces mobaraensis glutamine transaminage output, it is characterized in that adopting low temperature spray drying or lyophilize to make dry powder.
8. MgCl according to claim 7
2Coerce the method that improves streptomyces mobaraensis glutamine transaminage output, the condition that it is characterized in that described low temperature spray drying is 80 ℃ of inlet temperature, 60 ℃ of air outlet temperature 55-; Lyophilize adopts vacuum freeze drier dry, and the cold well temperature of Freeze Drying Equipment is-60 ℃, and vacuum tightness is 0.05MPa.
9. MgCl according to claim 3
2Coerce the method that improves streptomyces mobaraensis glutamine transaminage output, it is characterized in that by making the pharmaceutical grade zymin through freeze-drying again after ultrafiltration, gel chromatography, ion exchange chromatography, dialysis.
10. MgCl according to claim 9
2Coerce the method that improves streptomyces mobaraensis glutamine transaminage output, it is characterized in that the specific requirement of described preparation pharmaceutical grade zymin is as follows:
(1) ultrafiltration:
Above-mentioned acellular enzyme liquid is carried out ultrafiltration, and the ultrafiltration condition is the ultra-filtration membrane of 30kDa for adopting the molecular weight that dams, with 1/5 of acellular enzyme liquid simmer down to original volume;
(2) gel chromatography:
Concentrated solution is joined in the Sephadex G75 chromatography column of using in advance 50 mmol/L, pH 6.0 phosphate buffered saline buffer balances, and carry out wash-out with same buffer, flow velocity is 0.25 mL/min, detects wavelength for 280nm, collects activated part;
(3) ion exchange chromatography:
Whole active parts of collecting are joined in the SP sepharose High Performance chromatography column of using in advance 50 mmol/L, pH 6.0 phosphate buffered saline buffer balances, carry out linear gradient elution with the same buffer that contains 0-0.25mol/L sodium-chlor, flow velocity is 3 mL/min, detect wavelength for 280nm, collect active part.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107384820A (en) * | 2017-07-25 | 2017-11-24 | 华东理工大学 | One plant of glutamine transaminage high yield mutagenic strain and its application |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100497632C (en) * | 1999-09-30 | 2009-06-10 | 味之素株式会社 | Process for producing transglutaminase |
| CN100569943C (en) * | 2007-10-31 | 2009-12-16 | 天津大学 | Beta-mannase composite preparations and preparation method thereof |
-
2013
- 2013-03-07 CN CN2013100720243A patent/CN103146661A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100497632C (en) * | 1999-09-30 | 2009-06-10 | 味之素株式会社 | Process for producing transglutaminase |
| CN100569943C (en) * | 2007-10-31 | 2009-12-16 | 天津大学 | Beta-mannase composite preparations and preparation method thereof |
Non-Patent Citations (8)
| Title |
|---|
| LILI ZHANG ET AL.: "Enhancement of transglutaminase production in Streptomyces mobaraensis as achieved by treatment with excessive MgCl2", 《APPL MICROBIOL BIOTECHNOL》 * |
| LILI ZHANG ET AL.: "Enhancement of transglutaminase production in Streptomyces mobaraensis DSM 40587 by non-nutritional stress conditions: Effects of heat shock, alcohols, and salt treatments", 《KOREAN J. CHEM. ENG.》 * |
| 宋林等编著: "《生物技术学导论》", 31 December 2006, 北京市：中国人民大学出版社 * |
| 张莉丽: "MgCl2胁迫调控茂原链霉菌谷氨酰胺转氨酶合成及酶应用特性研究", 《万方数据知识服务平台》 * |
| 日本发酵工程学会编: "《微生物工程的基础和应用》", 30 September 1988, 轻工业出版社 * |
| 葛绍荣等: "《发酵工程原理与实践》", 31 August 2011, 华东理工大学出版社 * |
| 韩洛川编: "《生物技术学导论》", 31 December 2006, article "酶的分离纯化", pages: 110-113 * |
| 颜真，张英起主编: "《蛋白质研究技术》", 31 December 2007, 第四军医大学出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107384820A (en) * | 2017-07-25 | 2017-11-24 | 华东理工大学 | One plant of glutamine transaminage high yield mutagenic strain and its application |
| CN107384820B (en) * | 2017-07-25 | 2020-04-07 | 华东理工大学 | Glutamine transaminase high-yield mutant strain and application thereof |
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Application publication date: 20130612 |