CN1769439A - Solid state production technology for thermally-stabilised cellulase of feed use - Google Patents
Solid state production technology for thermally-stabilised cellulase of feed use Download PDFInfo
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- CN1769439A CN1769439A CNA2005100447497A CN200510044749A CN1769439A CN 1769439 A CN1769439 A CN 1769439A CN A2005100447497 A CNA2005100447497 A CN A2005100447497A CN 200510044749 A CN200510044749 A CN 200510044749A CN 1769439 A CN1769439 A CN 1769439A
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Abstract
A solid state production technique of cellulose for feeding with heat stability, belongs to feed microbe-additive skill field, which is characterized in that 1.Chaetomium is cultured on the inclined surface of the test tube at 45-55DEG C for 100-150hr; the seeds are cultured in conical flask at 45-55DEG C for 150- 180hr; leavens are cultured at 45-55DEG C, relative humidity at 83-88%, on the condition of natural draft until the surface of batch layer becomes greige. Culture chaetomin in solid state, and the culture medium is bran 100%, the ratio of material to water is 1: 1.1- 1.3, cool the system to 50-55DEG C after disinfection to seed follwing the quantity of 1%, ster stably. Keep the temperature at 40-45DEG C for 24hr and culture statically. The batch layer begins to appear white hypha after 24-30hr, and increase the temperature to 48-52DEG C after 48-54hr. The batch layer begins to agglomerate when the temperature is increased to 54-56DEG C, and we start to vent discontineously. Turn the leavens up one time when the time is 70- 72hr, vent continuously, draft cool air if the temperature is higher than 56DEG C to keep the temperature of the material to be 48-52DEG C. The batch layer begins to become alutaceous when the time is 90-120hr, stop venting and culturing. Dry pneumatic and stop when the water retentivity reaches 10.0-12.0%, analyze the activity of cellulose by CMC, the enzymatic activity per every gram production is 1500-3000U. 2. The heat stability of productions: handle the sample in baking box at the temperature of 60DEG C for 60 minutes, the loss rate of enzymatic activity is smaller than 3%, while at the temperature of 100DEG C for 60 minutes, the loss rate will reduce to smaller than 10%. The best temperature of said cellulose is 50-60DEG C. Solid state production technique has the advantages of simple craft, low cost, and the production thereby with good heat ability of enzymatic activity.
Description
Technical field
The present invention relates to a kind of solid state production technology of thermally-stabilised cellulase of feed use, belong to feed microbe additive technology field.
Background technology
Because cellulase belongs to biologically active substance, envrionment conditions to external world influence sensitivity, easier inactivation especially is heated.At present, the feeding cellulase product is all thermo-labile, and the loss of activity that especially is heated in the feed granulating process is huge, causes that actual use cost is too high, effect is not ideal enough.The feeding cellulase production technology mainly is divided into two kinds, i.e. liquid state fermentation production and solid state fermentation production, and liquid state fermentation production technology unit enzyme is lived lower, and product needed is separated, extracts, is concentrated, complex process, and also cost is higher; Solid-state fermentation technology unit's enzyme is lived higher, and product technology is simple, and cost is low, is suitable for producing feeding crude zyme preparation.The production of existing cellulase is mainly mould by mesophilous fungi wood, aspergillus, mould, the feeding cellulase poor heat stability that they produce, cost is higher, and production application is very limited, and the thermostability that therefore improves feeding cellulase has huge using value.Study a kind of technology of utilizing novel bacterial to produce feeding cellulase, thermo-labile, the defectives such as cost is high, complex process that solve present product existence become the major subjects of art technology tackling key problem.
Summary of the invention
The solid state production technology that the purpose of this invention is to provide a kind of thermally-stabilised cellulase of feed use, utilize the thermophilus strain chaetomium (C.thermophile) of high enzyme characteristic alive to produce heat stable feed Mierocrystalline cellulose crude zyme preparation by solid-state fermentation technology, improve to greatest extent enzyme to thermostability, and reduce production costs, and technology is simple, thereby overcomes the prior art above shortcomings.
The objective of the invention is to realize with following technical scheme:
Chaetomium test tube slant substratum is a solid PDA substratum, cultivates 100～150hr for 45～55 ℃.Triangular flask seed culture medium (wheat bran 100%, material-water ratio 1: 1.0～1.2) is cultivated 150～180hr for 45～55 ℃ then, middle button bottle 2 times.Plant bent substratum (wheat bran 100%, material-water ratio 1: 1.1～1.3), 45～55 ℃, natural ventilation is cultivated under the condition of relative humidity 83～88%, if temperature surpasses 55 ℃, then turns over Qu Jiangwen, after bed of material surface becomes beige behind 130～150hr, stop to cultivate, standby.
Chaetomium solid deep ventilation is cultivated, substratum (wheat bran 100%, material-water ratio 1: 1.1～1.3), and inoculation during sterilization postcooling to 50～55 ℃ by the inoculation of 1% inoculum size, is mixed thoroughly.Make temperature maintenance at 40～45 ℃ in the 24hr, static cultivation.24～30hr bed of material begins to occur white hypha, can be raised to 48～52 ℃ behind 48～54hr.When temperature was raised to 54～56 ℃, the bed of material began caking, carries out intermittent aeration.Turn over Qu Yici during 70～72hr, carry out continuous ventilating, when being higher than 56 ℃ as if temperature, logical freezing air makes the material temperature maintain 48～52 ℃.90～120hr bed of material begins thin out brown, stops to ventilate, and cultivates and finishes.Cultivate and finish, the material that ferments is shifted out the ventilation pond, carry out air stream drying,, adopt the CMC method to analyze the vigor of cellulase, every gram product enzyme 1500～3000U alive to moisture 10.0～12.0% o'clock end.
Product passes through 40 order sub-sieves through pulverizing more than 80%.Product is to heat stability test: sample is in 60 ℃ of oven 60 minutes, and enzyme is lived rate of loss less than 3%, 100 ℃ of oven 60 minutes, and enzyme is lived rate of loss less than 10%.50～60 ℃ of this cellulase the best use of temperature.
Feeding cellulase solid state production technology of the present invention has the advantage that technology is simple, cost is low, has the enzyme thermally-stabilised good advantage of living with the product of technological method production of the present invention.
Below in conjunction with embodiment the present invention is described in further details:
Embodiment 1: chaetomium test tube slant substratum is a solid PDA substratum, cultivates 150hr for 45 ℃.Triangular flask seed culture medium (wheat bran 100%, material-water ratio 1: 1.0) is cultivated 180hr for 45 ℃ then, middle button bottle 2 times.Plant bent substratum (wheat bran 100%, material-water ratio 1: 1.1), 45 ℃, natural ventilation is cultivated under the condition of relative humidity 83%, after bed of material surface becomes beige behind the 130hr, stops to cultivate, and is standby.
Chaetomium solid deep ventilation is cultivated, and substratum (wheat bran 100%, material-water ratio 1: 1.), by the inoculation of 1% inoculum size, mix thoroughly by inoculation during sterilization postcooling to 50 ℃.Make temperature maintenance at 40 ℃ in the 24hr, static cultivation.The 28hr bed of material begins to occur white hypha, can be raised to 50 ℃ behind the 52hr.When temperature was raised to 55 ℃, the bed of material began caking, carries out intermittent aeration.Turn over Qu Yici during 70hr, carry out continuous ventilating, make the material temperature maintain 48～52 ℃.The 110hr bed of material begins thin out brown, stops to ventilate, and cultivates and finishes.Cultivate end, the fermentation material is shifted out the ventilation pond, carry out air stream drying, moisture 11.2% adopts the CMC method to analyze the vigor of cellulase, every gram product enzyme 2600U alive.Product is through pulverizing, and 89% by 40 order sub-sieves.
Product is to heat stability test:
The enzyme powder is carried out Temperature Treatment, choose 7 temperature altogether: 40 ℃, 50 ℃, 60 ℃, 70 ℃, 80 ℃, 90 ℃, 100 ℃, and compare with former state.The enzyme powder: take by weighing 2.5 gram enzyme powder, its water content is 11.2%, places 90mm plate lid, and nominal is got 14 parts, puts in the baking oven and handles 1 hour, and each temperature is all done parallel, and the result is as follows:
| Temperature (℃) | CK | 40 | 50 | 60 | 70 | 80 | 90 | 100 |
| Enzyme (U) alive | 2600 | 2642 | 2760 | 2744 | 2636 | 2470 | 2392 | 2366 |
| Rate of loss (%) | 0 | 0 | 0 | 0 | 0 | 5 | 8 | 9 |
Embodiment 2: chaetomium test tube slant substratum is a solid PDA substratum, cultivates 100hr for 55 ℃.Triangular flask seed culture medium (wheat bran 100%, material-water ratio 1: 1.2) is cultivated 150hr for 55 ℃ then, middle button bottle 2 times.Plant bent substratum (wheat bran 100%, material-water ratio 1: 1.3), 55 ℃, natural ventilation is cultivated under the condition of relative humidity 88%, after bed of material surface becomes beige behind the 150hr, stops to cultivate, and is standby.
Chaetomium solid deep ventilation is cultivated, and inoculation when substratum (wheat bran 100%, material-water ratio 1: 1.3), sterilization postcooling to 55 ℃ by the inoculation of 1% inoculum size, is mixed thoroughly.Make temperature maintenance at 45 ℃ in the 24hr, static cultivation.The 25hr bed of material begins to occur white hypha, can be raised to 52 ℃ behind the 50hr.When temperature was raised to 56 ℃, the bed of material began caking, carries out intermittent aeration.Turn over Qu Yici during 72hr, make the material temperature maintain 48～52 ℃.The 95hr bed of material begins thin out brown, stops to ventilate, and cultivates and finishes.Cultivate end, the fermentation material is shifted out the ventilation pond, carry out air stream drying, moisture 10.0% adopts the CMC method to analyze the vigor of cellulase, every gram product enzyme 2300U alive.Product is through pulverizing, and 85% by 40 order sub-sieves.
Product is to heat stability test:
The enzyme powder is carried out Temperature Treatment, choose 7 temperature altogether: 40 ℃, 50 ℃, 60 ℃, 70 ℃, 80 ℃, 90 ℃, 100 ℃, and compare with former state.The enzyme powder: take by weighing 2.5 gram enzyme powder, its water content is 10.0%, places 90mm plate lid, and nominal is got 14 parts, puts in the baking oven and handles 1 hour, and each temperature is all done parallel, and the result is as follows:
| Temperature (℃) | CK | 40 | 50 | 60 | 70 | 80 | 90 | 100 |
| Enzyme (U) alive | 2500 | 2522 | 2560 | 2544 | 2486 | 2420 | 2302 | 2264 |
| Rate of loss (%) | 0 | 0 | 0 | 0 | 1 | 3 | 8 | 9 |
Claims (2)
- A kind of solid state production technology of thermally-stabilised cellulase of feed use is characterized in that:1,100～150hr is cultivated for 45～55 ℃ in the chaetomium test tube slant.The triangular flask seed is cultivated 150～180hr for 45～55 ℃.Plant bent 45～55 ℃, natural ventilation is cultivated under the condition of relative humidity 83～88%, after bed of material surface becomes beige, stops to cultivate.The solid-state cultivation of chaetomium, substratum are wheat bran 100%, material-water ratio 1: 1.1～1.3, and inoculation during sterilization postcooling to 50～55 ℃ by the inoculation of 1% inoculum size, is mixed thoroughly.Make temperature maintenance at 40～45 ℃ in the 24hr, static cultivation.24～30hr bed of material begins to occur white hypha, can be raised to 48～52 ℃ behind 48～54hr.When temperature was raised to 54～56 ℃, the bed of material began caking, carries out intermittent aeration.Turn over Qu Yici during 70～72hr, carry out continuous ventilating, when being higher than 56 ℃ as if temperature, logical freezing air makes the material temperature maintain 48～52 ℃.90～120hr bed of material begins thin out brown, stops to ventilate, and cultivates and finishes.Carry out air stream drying,, adopt the CMC method to analyze the vigor of cellulase, every gram product enzyme 1500～3000U alive to moisture 10.0～12.0% o'clock end.
- 2, product is to thermostability: sample is in 60 ℃ of oven 60 minutes, and enzyme is lived rate of loss less than 3%, 100 ℃ of oven 60 minutes, and enzyme is lived rate of loss less than 10%.50～60 ℃ of this cellulase the best use of temperature.
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| CNA2005100447497A CN1769439A (en) | 2005-09-22 | 2005-09-22 | Solid state production technology for thermally-stabilised cellulase of feed use |
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| CNA2005100447497A CN1769439A (en) | 2005-09-22 | 2005-09-22 | Solid state production technology for thermally-stabilised cellulase of feed use |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102771630A (en) * | 2012-06-07 | 2012-11-14 | 宁夏伊品生物科技股份有限公司 | Fermentation and coating of lysine and cellulose |
| CN102894190A (en) * | 2012-06-07 | 2013-01-30 | 宁夏伊品生物科技股份有限公司 | Fermentation and coating of lysine and compound enzyme |
| CN103343111A (en) * | 2013-07-05 | 2013-10-09 | 山东尤特尔生物科技有限公司 | Thermophilic chaetomium cellulase, as well as preparation method and application thereof |
-
2005
- 2005-09-22 CN CNA2005100447497A patent/CN1769439A/en active Pending
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102771630A (en) * | 2012-06-07 | 2012-11-14 | 宁夏伊品生物科技股份有限公司 | Fermentation and coating of lysine and cellulose |
| CN102894190A (en) * | 2012-06-07 | 2013-01-30 | 宁夏伊品生物科技股份有限公司 | Fermentation and coating of lysine and compound enzyme |
| CN102771630B (en) * | 2012-06-07 | 2013-10-16 | 宁夏伊品生物科技股份有限公司 | Fermentation and coating of lysine and cellulose |
| CN103355502A (en) * | 2012-06-07 | 2013-10-23 | 宁夏伊品生物科技股份有限公司 | Fermentation and coating of lysine and compound enzyme |
| CN103444983A (en) * | 2012-06-07 | 2013-12-18 | 宁夏伊品生物科技股份有限公司 | Lysine and cellulase fermented and coated feed |
| CN103355502B (en) * | 2012-06-07 | 2015-11-18 | 宁夏伊品生物科技股份有限公司 | The fermentation of Methionin, prozyme and bag quilt |
| CN103444983B (en) * | 2012-06-07 | 2015-11-18 | 宁夏伊品生物科技股份有限公司 | The fermentation of Methionin, cellulase and bag are by feed |
| CN103343111A (en) * | 2013-07-05 | 2013-10-09 | 山东尤特尔生物科技有限公司 | Thermophilic chaetomium cellulase, as well as preparation method and application thereof |
| CN103343111B (en) * | 2013-07-05 | 2015-08-26 | 山东尤特尔生物科技有限公司 | A kind of mesophilic protein and its preparation method and application |
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