CN102660527A - Method for producing high-enzyme-activity pectate lyase by means of raw material pretreatment and feed fermentation - Google Patents

Method for producing high-enzyme-activity pectate lyase by means of raw material pretreatment and feed fermentation Download PDF

Info

Publication number
CN102660527A
CN102660527A CN2012101723155A CN201210172315A CN102660527A CN 102660527 A CN102660527 A CN 102660527A CN 2012101723155 A CN2012101723155 A CN 2012101723155A CN 201210172315 A CN201210172315 A CN 201210172315A CN 102660527 A CN102660527 A CN 102660527A
Authority
CN
China
Prior art keywords
fermentation
enzyme
starch
medium
alkaline pectase
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
Application number
CN2012101723155A
Other languages
Chinese (zh)
Other versions
CN102660527B (en
Inventor
邹谋勇
李雪芝
赵建
曲音波
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shandong University
Original Assignee
Shandong University
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Shandong University filed Critical Shandong University
Priority to CN 201210172315 priority Critical patent/CN102660527B/en
Publication of CN102660527A publication Critical patent/CN102660527A/en
Application granted granted Critical
Publication of CN102660527B publication Critical patent/CN102660527B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Abstract

The invention discloses a method for producing high-enzyme-activity pectate lyase by means of raw material pretreatment and feed fermentation. The method is implemented by liquefying treatment and feed-batch fermentation for starch in a feed culture medium. The method obviously increases unit volume enzyme activity and volume production efficiency of the pectate lyase. Due to enzyme liquefaction treatment to the starch in the feed culture medium, viscosity coefficients of liquid are greatly reduced, and stirring efficiency is improved. Logarithmic growth phase of fungi is prolonged by means of feed fermentation, and biomass liveweight is increased. Experiments show that fermentation coarse enzyme fluid of the pectate lyase with the poly galacturonic acid cracking enzyme activity of 531UmL-1 is obtained by the method, and the highest volume production efficiency reaches 13U(mL*h)-1. The method further has the advantages of low cost, short fermenting period, high equipment utilization rate, simple process and suitability for large-scale industrial production.

Description

Utilize raw materials pretreatment and fed-batch fermentation to produce the method for high enzyme alkaline pectase alive
Technical field
The present invention relates to a kind of live method of alkaline pectase of high enzyme of producing, relate in particular to and a kind ofly produce the method for high enzyme work alkaline pectase, belong to the enzyme preparing technical field through the starch in the supplemented medium being carried out liquefaction processing and fed-batch fermentation.
Background technology
Polygalacturonase (pectinases) is that one type of enzyme that can decompose vegetable cell wall fraction pectin substance is a general name; From is olation, can polygalacturonase be divided into pectin hydrolase and pectin lyase, the former mainly comprises protopectinase (PPase), Rohapect MPE (PE), oligomeric galactose aldehydic acid lytic enzyme, gathers methyl galacturonate enzyme (PMG), polygalacturonase (PG); The latter mainly comprises polygalacturonic acid lyase (PGL) and gathers methyl galacturonate lyase (PMGL).The normal alkaline pectase of claiming of people refers generally to the polygalacturonic acid lyase in the pectin lyase.Pectin lyase breaks off glycosidic link through trans-elimination cracking pectin polymer on the C-4 position, simultaneously at H atom of C-5 place cancellation.Thereby produce a Δ 4:5 unsaturated link(age).
At present for the existing a large amount of bibliographical information of the research of polygalacturonase; Wherein the long acid pectase of search time is mainly used in the extraction of pectin fraction and the clarification of fruit wine fruit juice; Alkaline pectase is at pulping bleaching; Application in the cotton fabric treatment process has obtained certain progress, and particularly the applied research on the coming unstuck of bast-fibre increases day by day.Raw ramie is a kind of sheet bar that is bonded together by various colloids, and single strand is difficult for separately, therefore must be to the raw ramie processing of coming unstuck before the combing spinning.The mode of coming unstuck for a long time mainly is a chemical Degumming, and its ultimate principle is to utilize stable different to remove the colloid composition in the raw ramie to alkali, mineral acid and oxygenant of Mierocrystalline cellulose and colloid composition in the raw ramie.The shortcoming of this method is that violent chemical treatment possibly cause flaxen fiber impaired, influences the fine quality of linen, consumes a large amount of industrial chemicals and heat energy simultaneously, causes serious environmental to pollute.In recent years, biological degumming has received more concern, and it mainly comprises microbial degumming, and enzyme comes unstuck and the biological-chemical combination degumming.Wherein enzyme comes unstuck by force because of having flexibility of operation, and process is controlled easily, is convenient to add advantages such as boil-off assistant and the emphasis that becomes research.
Aspect the production of alkaline pectase, but Japanese scholar Koki Horkoshi in reported first in 1972 alkaliphile secreted alkaline polygalacturonase, Junwei Cao, T.SaKai, Tohru Kobayashi etc. also in succession separation screening to the bacterial strain that produces alkaline pectase.China was studied alkaline pectase since the beginning of the nineties, and the generation bacterium of having reported comprises Erwinia, Alkaliphilic bacillus, subtilis, spiral shell spore bacterium etc., reached the research to zymologic property but research work mainly rests on bacterial screening; Aspect the fermentation research of alkaline pectase, rest on mostly and shake a bottle condition optimizing, in the initial optimization research of small-sized fermentation jar condition of enzyme production.Liu Xi etc. optimize the condition of enzyme production of high-production alkalescence pectinase subtilis, make enzyme activity bring up to 14.82U mL -1(optimization of alkaline pectase superior strain condition of enzyme production. biotechnology, 2008,18 (6): 71-74.).Jiang Hongju etc. optimize the substratum and the fermentation culture conditions of Paenibacillus polymyxa fermentation product alkaline pectase, make enzyme work reach 311U mL -1(research of alkaline pectase fermentation condition is produced in Paenibacillus polymyxa 20185 liquid state fermentations. and China brewages, and 2011,235:111-114.).Dong Yunzhou etc. adopt the temperature control strategy that alkaline pectase is carried out fermentation optimization in the small-sized fermentation jar, and the enzyme work that makes genus bacillus WSH03-09 produce alkaline pectase reaches 5.99U mL -1(fermentation of bacillus produces alkaline pectase temperature control strategy. use and the environmental organism journal 2005,11 (3): 359-362.).Liu Huijuan etc. have studied the influence that differing temps (32-41 ℃) is produced the alkaline pectase kinetic parameter to genus bacillus WSHB04-02 batch fermentation, can make the enzyme work of alkaline pectase reach 53.0U mL behind the temperature optimization -1, production intensity is 3315U (hL) -1(fermentation of bacillus is produced the temperature control strategy of alkaline pectase. process engineering journal, 2007,7 (4): 786-789.).Chinese patent ZL 03131952.1 discloses the bacillus pumilus that alkaline pectase is produced in a strain, and it is 20UmL that liquid submerged fermentation obtains the alkaline pectase enzyme activity -1Chinese patent ZL 200410065807.X discloses a kind of through the transfer rate volume dissolved oxygen coefficient (K of oxygen in fermented liquid LA) method of raising alkaline pectin enzyme activity, its enzyme activity has reached 40U mL -1But the alkaline pectin enzyme activity and the industrial requirement of above-mentioned report also have a segment distance.Li Zuming etc. to gram Lloyd's genus bacillus carried out ultraviolet mutagenesis breeding and solid state fermentation conditions optimization (breeding of high-production alkalescence pectinase bacterial strain and the research of fermentation condition thereof. industrial microorganism, 2008,38 (3): 27-31.); Chinese patent ZL 94105708.9, CN 101096650B, ZL 200410073818.2, CN 101235361A disclose the method that the different strains solid fermentation is produced alkaline pectase, because the enzyme activity determination method differs, enzymatic productivity can't direct visual comparison; Simultaneously also rarely seen its is applied to industrial production because solid fermentation apparatus requires high, complex process.ZL97109044 discloses the method for a strain Alkaliphilic bacillus and China grass degumming thereof, but since usually time long (16-24h), the low industrial applications that also is difficult to of pectinase activity.ZL01106844.2 discloses a strain alkaline pectin enzyme-producing bacteria CCTCC NO:M200038, and the gained enzyme liquid that will ferment is applied to China grass degumming, fails to be applied to suitability for industrialized production owing to its fermenting enzyme vigor is lower.Therefore further optimization production technology; Through zymotechnique control and optimization and improvement alkaline pectase enzyme activity and volume production efficiency; Reducing the production cost of alkaline pectase, is the key that technological development and engineering are amplified, and also is the prerequisite that promotes the alkaline pectase industrial applications.Particularly deep liquid fermentation process optimization and the control for high concentration substrate seems particularly important.The report of the alkaline pectase of high yield adopts solid fermentation (high concentration of substrate) more at present, but its follow-up separation and purification process is complicated, and the zymoprotein yield is lower, has increased production cost.And high concentration substrate liquid submerged fermentation production alkaline pectase exists stirring efficiency low, problems such as mass transfer effect difference.The fermented liquid concentration of substrate has obtained in the time using widely with the prolongation enzymatic production fed-batch fermentation technology reducing just, and it has played vital role to fermentation middle and later periods cell growth division, keeping of cell viability, can effectively improve perhaps stabilized enzyme protein yield.In the method involving that retrieves, about also not appearing in the newspapers through the method for raw material being carried out pre-treatment and fed-batch fermentation production alkaline pectase.
Summary of the invention
In prior art, the alkaline pectin enzyme activity that liquid fermenting produces is low, and cost is high, is not suitable for the deficiency of suitability for industrialized production, the purpose of this invention is to provide a kind of method of utilizing raw materials pretreatment and fed-batch fermentation to produce high enzyme alkaline pectase alive.
The method of utilizing raw materials pretreatment and fed-batch fermentation to produce high enzyme alkaline pectase alive of the present invention; Be to improve alkaline pectase enzyme activity and production efficiency thereof through the starch in the supplemented medium being carried out liquefaction processing and fed-batch fermentation, its step is following:
(1) seed culture: under aseptic condition, activated inclined plane bacterial classification subtilis CCTCC NO:M200038, and be seeded to and carry out seed culture in the 50ml seed culture medium, 7-9h is cultivated in 36-37 ℃ of shaking table concussion;
(2) fermentation just: seed culture fluid is seeded to the 7.5L fermentor tank that fermention medium is housed, carries out fermentation culture, fermentor tank liquid amount 3-4L, inoculum size 10%, temperature 33-34 ℃, air flow 2.5-4.5L min -1
(3) the starch pre-treatment in the supplemented medium: warm glycase carries out liquefaction processing to the starch in the supplemented medium in using, and enzyme concentration is 20-45U g -1, liquefying-point is 45-65 ℃, liquefying time is 20-40min;
(4) fed-batch fermentation: just ferment to 13-19h, add pretreated supplemented medium by first fermention medium volume 10-25%, continue fermentation, temperature is 33-34 ℃, air flow 2.5-4.5L min -1, fermentation period is 36-48h;
(5) crude enzyme liquid preparation: after fermentation finished, with the centrifugal 10min of fermented liquid 12000rpm, supernatant was the crude enzyme liquid that contains alkaline pectase;
Above-mentioned seed culture medium is formed: glucose 8-13g L -1, yeast powder 3-6g L -1, peptone 3-6g L -1, NaCl 3-6g L -1, K 2HPO 48-12g L -1, pH 8.0;
Above-mentioned fermention medium is formed: starch 15-25g L -1, wheat bran 35-45g L -1, (NH 4) 2SO 41-2g L -1, MgSO 47H 2O 0.5-1.5g L -1
Above-mentioned supplemented medium is formed: starch 110-135g L -1, wheat bran 8-13g L -1, (NH 4) 2SO 46-14g L -1, MgSO 47H 2O 5-8g L -1
Above-mentioned utilization utilizes raw materials pretreatment and fed-batch fermentation to produce in the method for high enzyme alkaline pectase alive:
The preferred 115-130g L of starch concentration in the said supplemented medium -1
The preferred 25-40U g of warm glycase enzyme concentration during step (3) is said -1, the preferred 50-60 of liquefying-point ℃, the preferred 25-35min of liquefying time.
Step (4) is said to be added after the pre-treatment time of supplemented medium and is preferably and just ferments to 14-18h; Additional amount is preferably the 10-20% of fermention medium volume just, the optimum 20% disposable supplemented medium of adding by first fermention medium volume.
The on-line parameter that relates in the aforesaid method is measured
Dissolved oxygen (DO), pH, temperature adopt the on-line automatic control of electrode, and DO is relative dissolved oxygen level, that is: the 30min that before substratum is not inoculated, ventilates makes the substratum dissolved oxygen level that reaches capacity, this moment DO to demarcate be 100%, it is 0% that the saturated sodium sulfite dissolved oxygen is demarcated.
The offline parameter that relates in the aforesaid method is measured
Living weight: get fermented liquid 1mL (with the centrifugal back of fermented liquid supernatant as contrast) adds 1M in tube comparison tubes perchloric acid 1mL; Behind the boiling water bath 20min; Go to the centrifugal 10min of 10000rpm in the centrifuge tube; Get supernatant 1ml constant volume to 10mL, measure its light absorption value in ultraviolet spectrophotometer 260nm place and measure light absorption value, reference standard curve calculation dry cell weight.The standard curve making method is: dilution obtains the concentration gradient thalline, measures OD respectively 260, and corresponding weighing dry cell weight, be the X axle with the dried cell weight, OD 260Be Y axle drawing standard curve.
Alkaline pectase (PGL) activity: get the 5mL fermented liquid; With 1200rpm; 4 ℃ of centrifugal 10min get supernatant and get the polygalacturonic acid solution that 20 μ L add 2mL 0.2% (w/v) after with pH 9.6Gly-NaOH damping fluid dilution certain multiple and (with pH 9.6, contain 0.44mM CaCl 2, the configuration of 0.05M Gly-NaOH damping fluid), 45 ℃ of reaction 15min with 3mL0.03M phosphoric acid solution termination reaction, measure its light absorption value in ultraviolet spectrophotometer 235nm place.
A standard enzyme unit of activity (IU) is defined as: the unit time (1min) makes the polygalacturonic acid cleavage produce the enzyme amount of 1 μ mol unsaturated polyester galacturonic acid.The molar absorptivity of wherein unsaturated galacturonic acid at wavelength 235nm place is 4600 (M).
Beneficial effect of the present invention:
Through adopting glycase that the high density starch in the supplemented medium is carried out liquefaction processing; Effectively reduce the viscosity factor of feed supplement liquid; Improved the flowability and the oxygen carrying capacity of feed liquid, reduced fermentor tank and stirred resistance, for pump flow feeding substratum provides possibility; Improve dissolving sugar concentration in the substrate simultaneously, make the nutrition of cell rapidly obtain, division growth has improved the growth velocity of subtilis fast.Prolong the thalli growth time through fed-batch fermentation, improve living weight, prolong effective integration time of alkaline pectase; Make the unit enzyme activity of alkaline pectase reach 531U mL -1, volume production efficiency reaches 13U (mlh) -1(not appearing in the newspapers both at home and abroad).This method has shortened fermentation period greatly, thereby has improved the fermentor tank utilising efficiency, has reduced production cost; Operation is simple for this method, for suitability for industrialized production has been established solid basis.The alkaline pectase of fermentative prodn just enzyme liquid can be used for processings of coming unstuck of fibrilia raw material, can effectively shorten even saves the chemical Degumming operation, improves the quality of flaxen fiber product, and the minimizing waste liquid is to the pollution of environment.
Description of drawings
Fig. 1 is the influence of fed-batch fermentation to the alkaline pectin enzyme activity.
Fig. 2 is that the influence to the alkaline pectin enzyme activity is handled in the liquefaction of starch material glycase in the supplemented medium.
Embodiment
Embodiment 1
(applicant was preserved in Chinese typical culture center (CCTCC) to subtilis CCTCC NO:M200038 on November 20th, 2000; Related patent U.S. Patent No. is seen ZL01106844.2) through slant activation; Provoke lawn be seeded to be equipped with seed culture medium shake the bottle cultivate; Culture condition is: 300ml triangular flask liquid amount is 50ml, 37 ℃ of leavening temperatures, and 8h is cultivated in the 200rpm concussion.With the first fermention medium of inoculum size 10% inoculation, temperature 33-34 ℃, air flow 2.5-4.5L min -1Warm glycase carries out liquefaction processing to the starch in the supplemented medium in using, and enzyme concentration is 25U g -1, liquefying-point is 60 ℃, liquefying time is 35min.Just ferment to 14h, add the later supplemented medium of pre-treatment by 20% of first fermention medium volume, controlled temperature 33-34 ℃, air flow 2.5-4.5L min -1Fermentation period is 36-48h.After fermentation finished, with the centrifugal 10min of fermented liquid 12000rpm, supernatant was the crude enzyme liquid that contains alkaline pectase; Through measuring, the high alkalinity pectinase activity of said crude enzyme liquid is 465U mL -1Fermenting process is seen Fig. 1.Wherein culture medium prescription is following:
Seed culture medium is formed: glucose 8g L -1, yeast powder 3g L -1, peptone 3g L -1, NaCl 3g L -1, K 2HPO 48g L -1, pH 8.0;
Fermention medium is formed: starch 15g L -1, wheat bran 35g L -1, (NHz 4) 2SO 41g L -1, MgSO 47H 2O 0.5gL -1
Supplemented medium is formed: starch 115g L -1, wheat bran 8g L -1, (NH 4) 2SO 46g L -1, MgSO 47H 2O 5g L -1
Embodiment 2
Fermentation process, condition are except that following change, and all the other are with embodiment 1.In the preprocessing process, middle temperature glycase enzyme concentration is 32.5U g -1, liquefying-point is 55 ℃, liquefying time is 30min.Just ferment to the 16h feed supplement.Fermentation period is 36-48h, and the high alkalinity pectinase activity of the crude enzyme liquid of acquisition is 531U mL -1Fermenting process is seen Fig. 1.Wherein culture medium prescription is following:
Seed culture medium is formed: glucose 11g L -1, yeast powder 4,5g L -1, peptone 4.5g L -1, NaCl 4.5g L -1, K 2HPO 410g L -1, pH 8.0;
Fermention medium is formed: starch 20g L -1, wheat bran 40g L -1, (NH 4) 2SO 41.5g L -1, MgSO 47H 2O 1gL -1
Supplemented medium is formed: starch 122.5g L -1, wheat bran 10.5g L -1, (NH 4) 2SO 410g L -1, MgSO 47H 2O6.5g L -1
Embodiment 3
Fermentation process, condition are except that following change, and all the other are with embodiment 1.In the preprocessing process, middle temperature glycase enzyme concentration is 40U g -1, liquefying-point is 50 ℃, liquefying time is 25min.Just ferment to the 18h feed supplement.Fermentation period is 36-48h, and the final alkaline pectin enzyme activity of the crude enzyme liquid of acquisition is 479UmL -1Fermenting process is seen Fig. 1.Wherein culture medium prescription is following:
Seed culture medium is formed: glucose 13g L -1, yeast powder 6g L -1, peptone 6g L -1, NaCl 6g L -1, K 2HPO 412g L -1, pH 8.0;
Above-mentioned fermention medium is formed: starch 25g L -1, wheat bran 45g L -1, (NH 4) 2SO 42g L -1, MgSO 47H 2O 1.5g L -1
Above-mentioned supplemented medium is formed: starch 130g L -1, wheat bran 13g L -1, (NH 4) 2SO 414g L -1, MgSO 47H 2O8g L -1
Embodiment 4
Fermention medium and supplemented medium are mixed, carry out batch fermentation (being not carry out feed supplement in the process), seed activation, cultivation, inoculum size, leavening temperature and air flow are identical with embodiment 1.Fermentation 48h, the final alkaline pectin enzyme activity of the crude enzyme liquid of acquisition is 203U mL -1Fermenting process is seen Fig. 1.Compare with embodiment 1,2,3, the crude enzyme liquid alkaline pectase enzyme activity that present embodiment obtains is obviously lower.
Embodiment 5
Seed activation, cultivation and inoculum size are carried out the fed-batch fermentation experiment, 122.5g L in the experimental group supplemented medium with embodiment 1 on the 500mL triangular flask -1High density starch with in warm glycase carry out liquefaction processing, enzyme concentration is 45U g -1, liquefying-point is 65 ℃, liquefying time is 40min.Handle as contrast not carry out glycase.This condition bottom fermentation 60h, the crude enzyme liquid alkaline pectase enzyme activity of acquisition reaches 347U ml -1Fermenting process is seen Fig. 2.
Embodiment 6
Fermentation process, condition are except that following change, and all the other are with embodiment 1, and starch concentration is 110g L in the supplemented medium -1, middle temperature glycase enzyme concentration is 20U g -1, liquefying-point is 60 ℃, liquefying time is 20min.Just ferment to the 16h feed supplement.Fermentation period 36-48h, the high alkalinity pectinase activity of the crude enzyme liquid of acquisition is 437U mL -1
Embodiment 7
Fermentation process, condition are except that following change, and all the other are with embodiment 1, and starch concentration is 135g L in the supplemented medium -1, middle temperature glycase enzyme concentration is 45U g -1, liquefying-point is 45 ℃, liquefying time is 30min.Just ferment to the 16h feed supplement, add supplemented medium according to 25% first fermention medium volume.Fermentation period 36-48h, the high alkalinity pectinase activity of the crude enzyme liquid of acquisition is 421U mL -1
Embodiment 8
Fermentation process, condition are except that following change, and all the other are with embodiment 1, just ferment to 14h to add supplemented medium by 10% of first fermention medium volume, and fermentation period 36-48h, the high alkalinity pectinase activity of the crude enzyme liquid of acquisition is 340U mL -1

Claims (4)

1. one kind is utilized raw materials pretreatment and fed-batch fermentation to produce the live method of alkaline pectase of high enzyme, and step is:
(1) seed culture: under aseptic condition, activated inclined plane bacterial classification subtilis CCTCC NO:M200038, and be seeded to and carry out seed culture in the 50ml seed culture medium, 7-9h is cultivated in 36-37 ℃ of shaking table concussion;
(2) fermentation just: seed culture fluid is seeded to the 7.5L fermentor tank that fermention medium is housed, carries out fermentation culture, fermentor tank liquid amount 3-4L, inoculum size 10%, temperature 33-34 ℃, air flow 2.5-4.5L min -1
(3) the starch pre-treatment in the supplemented medium: warm glycase carries out liquefaction processing to the starch in the supplemented medium in using, and enzyme concentration is 20-45U g -1, liquefying-point is 45-65 ℃, liquefying time is 20-40min;
(4) fed-batch fermentation: just ferment to 13-19h, add pretreated supplemented medium by first fermention medium volume 10-25%, continue fermentation, temperature is 33-34 ℃, air flow 2.5-4.5L min -1, fermentation period is 36-48h;
(5) crude enzyme liquid preparation: after fermentation finished, with the centrifugal 10min of fermented liquid 12000rpm, supernatant was the crude enzyme liquid that contains alkaline pectase;
Above-mentioned seed culture medium is formed: glucose 8-13g L -1, yeast powder 3-6g L -1, peptone 3-6g L -1, NaCl 3-6g L -1, K 2HPO 48-12g L -1, pH 8.0;
Above-mentioned fermention medium is formed: starch 15-25g L -1, wheat bran 35-45g L -1, (NH 4) 2SO 41-2g L -1, MgSO 47H 2O 0.5-1.5g L -1
Above-mentioned supplemented medium is formed: starch 110-135g L -1, wheat bran 8-13g L -1, (NH 4) 2SO 46-14g L -1, MgSO 47H 2O 5-8g L -1
2. utilize raw materials pretreatment and fed-batch fermentation to produce the method for high enzyme alkaline pectase alive according to claim 1, it is characterized in that the starch concentration in the said supplemented medium is 115-130g L -1
3. utilize raw materials pretreatment and fed-batch fermentation to produce the method for high enzyme alkaline pectase alive according to claim 1, it is characterized in that warm diastatic enzyme concentration was 25-40U g during step (3) was said -1, liquefying-point is 50-60 ℃, liquefying time is 25-35min.
4. utilize raw materials pretreatment and fed-batch fermentation to produce the method for high enzyme alkaline pectase alive according to claim 1; It is characterized in that; The time that step (4) is said adds supplemented medium after the pre-treatment, additional amount was the 10-20% of first fermention medium volume in order just to ferment to 14-18h.
CN 201210172315 2012-05-30 2012-05-30 Method for producing high-enzyme-activity pectate lyase by means of raw material pretreatment and feed fermentation Active CN102660527B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN 201210172315 CN102660527B (en) 2012-05-30 2012-05-30 Method for producing high-enzyme-activity pectate lyase by means of raw material pretreatment and feed fermentation

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN 201210172315 CN102660527B (en) 2012-05-30 2012-05-30 Method for producing high-enzyme-activity pectate lyase by means of raw material pretreatment and feed fermentation

Publications (2)

Publication Number Publication Date
CN102660527A true CN102660527A (en) 2012-09-12
CN102660527B CN102660527B (en) 2013-07-03

Family

ID=46770107

Family Applications (1)

Application Number Title Priority Date Filing Date
CN 201210172315 Active CN102660527B (en) 2012-05-30 2012-05-30 Method for producing high-enzyme-activity pectate lyase by means of raw material pretreatment and feed fermentation

Country Status (1)

Country Link
CN (1) CN102660527B (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106434476A (en) * 2016-11-02 2017-02-22 青岛蔚蓝生物集团有限公司 High-yield strain for alkaline pectinase and application of high-yield strain

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0798377A2 (en) * 1996-03-29 1997-10-01 Kyowa Hakko Kogyo Kabushiki Kaisha Process for producing aspartase and l-aspartic acid
CN1330207A (en) * 2001-02-22 2002-01-09 湖南辰州矿业有限责任公司 Hydraulic prop system
CN101705252A (en) * 2009-12-16 2010-05-12 湘潭大学 Method for preparing alcohol from high-concentration raw dry sweet potato mash by using feed fermentation method

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0798377A2 (en) * 1996-03-29 1997-10-01 Kyowa Hakko Kogyo Kabushiki Kaisha Process for producing aspartase and l-aspartic acid
US5916782A (en) * 1996-03-29 1999-06-29 Kyowa Hakko Kogyo Co., Ltd. Process for producing aspartase and process for producing L-aspartic acid
CN1330207A (en) * 2001-02-22 2002-01-09 湖南辰州矿业有限责任公司 Hydraulic prop system
CN101705252A (en) * 2009-12-16 2010-05-12 湘潭大学 Method for preparing alcohol from high-concentration raw dry sweet potato mash by using feed fermentation method

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
李雪芝: "造纸工业用均知的选育和相关酶的研究及应用", 《中国优秀硕士论文全文数据库》, 15 December 2006 (2006-12-15) *
赵建: "两株淀粉酶产生菌的分类鉴定、酶学性质分析及一株果胶酶产生菌的分类鉴定", 《中国优秀硕士论文全文数据库》, 15 February 2009 (2009-02-15) *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106434476A (en) * 2016-11-02 2017-02-22 青岛蔚蓝生物集团有限公司 High-yield strain for alkaline pectinase and application of high-yield strain

Also Published As

Publication number Publication date
CN102660527B (en) 2013-07-03

Similar Documents

Publication Publication Date Title
Liu et al. Consolidated bioprocess for bioethanol production with alkali-pretreated sugarcane bagasse
CN103045566A (en) Method for producing cellulase by using induction and regulation of substrate
CN111349565B (en) Method for culturing chlorella pyrenoidosa with high biomass and high protein content
CN110205349A (en) A method of bacteria cellulose is prepared using rice bran hydrolyzate fermentation
CN107686854B (en) Method for degrading and modifying schizophyllan by utilizing endoprotease produced by schizophyllum commune fermentation system
CN103045484B (en) Penicillium strain producing cellulase and application in cellulose enzymatic hydrolysis thereof
CN101948883B (en) Method for fermentatively producing microbial oil by utilizing corn husk dregs as raw materials
CN103421851B (en) A kind of method preparing sugar and ethanol with sweet potato waste
CN106801075A (en) A kind of production method of rhamnolipid
CN103789360B (en) A kind of corn cob fiber element fermentation that utilizes is for the method for fumaric acid fermented liquid
JP2015519070A (en) Method for producing enzyme cocktail using liquid residue from biochemical conversion process of lignocellulosic material
CN103789354A (en) Method for preparing ethanol from cellulose-containing raw material
CN102234670B (en) Method for producing bacterial cellulose through solid state fermentation by using inert adsorption carrier
CN101173303B (en) Method for vapor-exploding stalk enzymolysis coupling ferment for hydrogen production by using immobilized cell
CN102660527B (en) Method for producing high-enzyme-activity pectate lyase by means of raw material pretreatment and feed fermentation
CN102660526B (en) Method for producing pectate lyase by two-section pH (potential of hydrogen) control
Zeng et al. Efficient production of scleroglucan by Sclerotium rolfsii and insights into molecular weight modification by high-pressure homogenization
CN107177637A (en) The methods and applications pre-processed using microorganism to maize straw hydrolysis and saccharification
CN102220244A (en) Hypocrea lixii strain and method for preparing dextran enzyme with the Hypocrea lixii strain
CN104160021A (en) Method for producing an enzyme cocktail using the solid residues from a process for biochemically converting of lignocellulosic materials
CN102286572A (en) Method for preparing fermentable sugar solution from straws
CN102876756B (en) Process for co-producing lactic acid with lower polyxylose
CN108013212A (en) A kind of method using manioc waste and its alcohol waste mash production feed addictive
CN102766656A (en) Method for cheaply preparing microbial flocculant by utilizing bagasse
CN102154249A (en) Culture method for producing high-activity beta-glucosidase

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