CN102250857A - Liquid fermentation technology capable to improve per unit of cellulose activity - Google Patents
Liquid fermentation technology capable to improve per unit of cellulose activity Download PDFInfo
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- CN102250857A CN102250857A CN2010101768833A CN201010176883A CN102250857A CN 102250857 A CN102250857 A CN 102250857A CN 2010101768833 A CN2010101768833 A CN 2010101768833A CN 201010176883 A CN201010176883 A CN 201010176883A CN 102250857 A CN102250857 A CN 102250857A
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Abstract
The invention discloses a liquid fermentation technology capable to improve per unit of cellulose activity, and belongs to the field of microbial fermentation. The fermentation technology adopts trichoderma reesei Rut C-30 as a cellulose fermentation strain, and is characterized by optimizing a proportion between microcrystalline cellulose and bran in a culture medium, and then optimizing factors of air volumes and fermentation times of a seeding tank and a fermentation tank. The factors have a very important influence on an enzymatic activity. The technology has the advantages of mature process, wide raw material source, low cost, excellent stability and good applicability for a large-scale industrialization production. An activity of fermented enzyme liquid produced by the technology can reach to 1200 u/g, and is above 25% higher than that of the domestic average fermentation level.
Description
Technical field: the invention belongs to microorganisms producing and make the field, the present invention relates to a kind of liquid fermentation process that improves the cellulase units activity, specifically be to optimize the fermention medium proportioning, and improve seeding tank and fermentor tank condition, thereby improve the plain enzymic activity of unit fiber.
Technical background:
Mierocrystalline cellulose is renewable natural resource the abundantest on the earth as the main polyose product of photosynthesis of plant.According to estimates, earth Mierocrystalline cellulose is about 4 * 10 by photosynthetic renewal amount every year
10T, the energy of being contained are about as much as the contained energy of oil of 6,400 hundred million t, and cellulosic recyclability is that the mineral energy such as oil are incomparable.Yet about 80% Mierocrystalline cellulose is not exploited at present, has very tempting prospect.
The present cellulase that studies show that is a class inducible enzyme, starch-containing in the wheat bran, Mierocrystalline cellulose, hemicellulose, organonitrogen and somatomedin, the abundant nutrition material that it contains can promote the growth of thalline, and contained Mierocrystalline cellulose and hemicellulose may have inducing action to enzyme synthetic.
Microcrystalline Cellulose is the more satisfactory cheap carbon source of fermentative production cellulase, and cellulase is also had inducing action preferably.Be accompanied by in the building-up process of cellulase and produce acid, and may have inducing action, thereby pH there is material impact to producing enzyme the synthetic of cellulase.But under high density carbon source condition, because the big volume production acid of carbon source metabolism thalline, nutrient solution pH sharply descends, thereby causes the stagnation of the inactivation and the thalli growth of enzyme, and is unfavorable for producing enzyme.This patent is on a large amount of bases, further optimized the proportioning of Microcrystalline Cellulose and wheat bran, the optimum concn of accurately obtaining them is: Microcrystalline Cellulose is 1.4%, wheat bran is 3.5%, and pH is controlled effectively, and makes it maintain the best enzyme district that produces all the time.
Substratum, carbon source: Microcrystalline Cellulose 1.4%, wheat bran 3.5%
Auxiliary material: soybean cake powder 0.8%-1%, KH
2PO
40.4%,
Peptone 0.2%, yeast powder 0.05%,
Behind the shake-flask culture 12h, change over to and carry out liquid submerged fermentation in seeding tank and the fermentor tank.With the above substratum fermentor tank of packing into of sizing mixing, first class seed pot 100L, fermentation time 30h, ventilation is 20m
3/ h; Second seed jar 1t, fermentation time 40h, ventilation is 29m
3/ h produces a jar 15t, and ventilation is 360m
3/ h finishes fermentation through 55h.But production process on-line monitoring pH if any unusually, keeps the pH value and gets final product near 5.5.Fermented liquid separates to remove residue through Plate Filtration or whizzer, is concentrated into 1/10 of raw material volume behind the filtrate collection, then the spray-dried operation of concentrated solution is made the high active cellulase powder.
The measuring method of cellulase activity: with 20mg Xinhua filter paper is substrate, the acetate buffer solution 0.9ml that adds pH4.8,0.1M, the enzyme liquid 0.1ml that adds suitable dilution again, 50 ℃ of reactions 60 minutes, with 3, the reducing sugar that 5-dinitrosalicylic acid method (DNS method) measure to generate, sugar with glucose as a standard, promptly to produce 1 μ g glucose be an enzyme activity unit to per minute.
Summary of the invention:
The invention discloses a kind of liquid fermentation process that improves the cellulase units activity.This method is specific as follows: substratum proportioning, carbon source: Microcrystalline Cellulose 1.4%, wheat bran 3.5%;
Auxiliary material: soybean cake powder 0.8%-1%, KH
2PO
40.4%, peptone 0.2%, yeast powder 0.05%.
Behind the shake-flask culture 12h, change over to and carry out liquid submerged fermentation in seeding tank and the fermentor tank.With the above substratum fermentor tank of packing into of sizing mixing, first class seed pot 100L, fermentation time 30h, ventilation is 20m
3/ h; Second seed jar 1t, fermentation time 40h, ventilation is 29m
3/ h produces a jar 15t, and ventilation is 360m
3/ h finishes fermentation through 55h.But production process on-line monitoring pH if any unusually, keeps the pH value and gets final product near 5.5.Fermenting enzyme liquid activity is considered oven dry loss about 1200u/g, utilize enzyme powder activity that this method produces at 10000u/g, surpasses offshore company's like product.
Below in conjunction with embodiment the present invention is further specified:
Embodiment 115t fermentation production process
1, a seeding tank ventilation is 20m
3/ h, second seed jar ventilation is 29m
3/ h, producing a jar ventilation is 360m
3/ h.
2, sky temperature 125 degree that disappears, 30 minutes; The temperature that disappears in fact 123 degree, 40 minutes time;
The pipeline that disappears is 0.3Mpa, 2 hours.
3, seeding tank pH nature; Tank pressure all is 0.05MPa;
4, constant volume 66L before a seeding tank disappears; The second seed jar preceding constant volume 650L that disappears; 10 tons of constant volumes (the seeding tank sterilization all is warmed up to 90 degree with adding to overlap earlier) before the production jar disappears
5, referential data: first class seed pot was cultivated 30 hours, enzyme 200-400u/ml alive
The secondary seed jar was cultivated 40 hours, and enzyme is lived more than the 400-600u/ml
Production jar cultivation is enzyme 1100-1300u/ml alive in the time of 55 hours
Description of drawings:
Figure 110 0L seeding tank synoptic diagram.
Fig. 2 production unit schema.
Claims (3)
1. a liquid fermentation process that improves the cellulase units activity adopts Microcrystalline Cellulose and wheat bran as raw material, adds soybean cake powder, KH
2PO
4, (NH
4) SO
4Make substratum Deng auxiliary material, after sterilising treatment, inoculate test tube and triangle shakes bottle, carry out cultivation of secondary seed jar and fermentation, it is characterized in that Microcrystalline Cellulose and wheat bran are as carbon source, as the cellulase fermentations bacterial strain, in secondary seed jar and fermentor tank, carry out liquid submerged fermentation with Richter scale wood enzyme Rut C-30.The fermentor tank coefficient is 66%.
2. according to claims 1 described substratum, it is characterized in that substratum proportioning the best is Microcrystalline Cellulose 1.4%, wheat bran 3.5%, soybean cake powder 1%, KH
2PO
40.4%, peptone 0.2%, yeast powder 0.05%, tap water is mixed with 100%, natural pH value.
3. according to carrying out liquid submerged fermentation in claims 1 described seeding tank and the fermentor tank, it is characterized in that first class seed pot 100L, fermentation time 30h, ventilation is 20m
3/ h; Second seed jar 1t, fermentation time 40h, ventilation is 29m
3/ h produces a jar 15t, fermentation time 55h, and ventilation is 360m
3/ h, after fermentation finishes after filtration, concentrate, the spraying drying operation makes the high active cellulase powder.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010101768833A CN102250857A (en) | 2010-05-17 | 2010-05-17 | Liquid fermentation technology capable to improve per unit of cellulose activity |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010101768833A CN102250857A (en) | 2010-05-17 | 2010-05-17 | Liquid fermentation technology capable to improve per unit of cellulose activity |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102250857A true CN102250857A (en) | 2011-11-23 |
Family
ID=44978428
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|---|---|---|---|
| CN2010101768833A Pending CN102250857A (en) | 2010-05-17 | 2010-05-17 | Liquid fermentation technology capable to improve per unit of cellulose activity |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102876590A (en) * | 2012-10-17 | 2013-01-16 | 广西大学 | Penicillium sp. mutant strain and application of penicillium sp. mutant strain to cellulase preparation |
| CN102994481A (en) * | 2012-12-05 | 2013-03-27 | 天津工业生物技术研究所 | Preparation method for compound enzyme system for high-efficiency degradation for lignocellulose and application thereof |
| CN105779301A (en) * | 2016-03-24 | 2016-07-20 | 山东大学 | Trichoderma reesei as well as culture method thereof and application thereof |
-
2010
- 2010-05-17 CN CN2010101768833A patent/CN102250857A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102876590A (en) * | 2012-10-17 | 2013-01-16 | 广西大学 | Penicillium sp. mutant strain and application of penicillium sp. mutant strain to cellulase preparation |
| CN102876590B (en) * | 2012-10-17 | 2013-11-13 | 广西大学 | Penicillium sp. mutant strain and application of penicillium sp. mutant strain to cellulase preparation |
| CN102994481A (en) * | 2012-12-05 | 2013-03-27 | 天津工业生物技术研究所 | Preparation method for compound enzyme system for high-efficiency degradation for lignocellulose and application thereof |
| CN105779301A (en) * | 2016-03-24 | 2016-07-20 | 山东大学 | Trichoderma reesei as well as culture method thereof and application thereof |
| CN105779301B (en) * | 2016-03-24 | 2019-07-16 | 山东大学 | One plant of trichoderma reesei and its cultural method and application |
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Application publication date: 20111123 |