CN101870965A - Production method of high enzymic activity instant enzyme preparation - Google Patents
Production method of high enzymic activity instant enzyme preparation Download PDFInfo
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- CN101870965A CN101870965A CN201010188024A CN201010188024A CN101870965A CN 101870965 A CN101870965 A CN 101870965A CN 201010188024 A CN201010188024 A CN 201010188024A CN 201010188024 A CN201010188024 A CN 201010188024A CN 101870965 A CN101870965 A CN 101870965A
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Abstract
The invention discloses a production method of a high enzymic activity instant enzyme preparation, which belongs to the field of biotechnology, and comprises the step of carrying out protein salting-out sedimentation and vacuum freezing drying on fermentation liquor after being separated and concentrated in the post-treatment process of the fermentation liquor after high-density fermentation. The method for producing the enzyme preparation can increase the stability of the enzyme and is convenient for goods transportation. The obtained enzyme preparation product has high enzymic activity which is more than 25 times higher than that of the fermentation liquor, the loss of active ingredients of the product is small, the original physical and chemical properties of the material can be maintained, the water content is less than 3%, the product has long shelf life and good stability, can be dissolved in water instantly, and the enzyme liquor after dissolution is clear with high clarity, and is more convenient to use.
Description
Technical field
The present invention relates to a kind of production method of high enzymic activity instant enzyme preparation, is specifically in the last handling process of fermented liquid, and the concentrated solution of enzyme is carried out albumen salt precipitation and vacuum lyophilization, belongs to biological technical field.
Background technology
Zymin industry is the knowledge-intensive high-tech industry, is bionic important economic entity.According to statistics, zymin market in the whole world increases year by year with the speed of annual 11% in recent years.With regard to the application of enzyme in each field, food and fodder industry consumption maximum account for 45% of gross sales (GS), and washing composition accounts for 32%, and textile industry accounts for 11%, and paper industry accounts for 7%, and chemical industry accounts for 4%.Foodstuffs industry accounts for zymin market 62% with enzyme during with 1985, and enzyme for detergent accounts for 33%, and the process hides textile industry accounts for 5% with enzyme and compares, and its obvious variation is that the enhancing of people to environmental consciousness reflected in rapid expansion in non-food product industrial enzymes field.At present zymin has been widely used in a lot of fields such as herding, food, the energy, weaving, washing composition, and Application Areas constantly enlarges, and the utilisation technology level improves constantly.21 century will be conceived to development of biology, and guiding is rebuild traditional industry, and cut down the consumption of energy, and administer environment.In the target of China's Eleventh Five-Year Plan, energy consumption reduces by 20% than Tenth Five-Year Plan Period, fields such as bioenergies such as development alcohol fuel, biofuel, biological environmental production, biological manufacturing, in the Application Areas of zymin, focus accounts for critical role in national economy but industry such as raw material and energy consumption are big, environmental pollution is serious papermaking, weaving, process hides, feed.
Enzyme is a kind of specific proteins with height catalytic activity.Industrial liquid enzyme formulation must concentrate and purify with convenient transportation and use.Traditional processing technology is fermentation, flocculation sediment, filtration, solvent extraction, vacuum-evaporation, drying, and its production process energy consumption height, enzyme deactivation rate height, yield are low.The molecular mass of zymin commonly used is between 10000 ~ 100000Dal, this scope is just within the scope that ultra-filtration technique is used, thereby people concentrate purification with membrane technique to enzymic fermentation liquid, and wherein the maximum of usefulness are micro-filtration, ultrafiltration, nanofiltration and reverse osmosis.Handling fermented liquid with membrane technique can remove with the purpose molecular weight of product according to the size of molecular weight of material and differ bigger all kinds of impurity, reduce the influence of temperature to the zymin quality, improve the fermented liquid quality, and in sepn process, not relating to phase transformation, non-secondary pollution has the function of microbial film enrichment method again, the carrying out that helps the subsequent process process, and improve product purity and yield, and reduce solvent-oil ratio, cut down the consumption of energy.Since easy to operate, compact construction, and maintenance cost is low, is easy to automatization, thereby membrane technique is applied in the last handling process of multiple leavened prod.
Existing zymin kind also can not satisfy the needs of application far away, and zymin industry is constantly being released novel enzyme preparation, compound enzymic preparation, high vigor and high purity specific enzymes preparation and satisfied different needs.But the influence of factors such as the stability of zymin is subject to transport, storage, and storage, transportation inconvenience.How under the situation that the zymologic property of zymin does not change, existing zymin height is concentrated, strengthen its stability, storage, transportation are simplified greatly, use more conveniently, a kind of ideal method solution is not also arranged so far.
Summary of the invention
At the deficiency that prior art exists, technical problem to be solved by this invention is that a kind of production method of high enzymic activity instant enzyme preparation is provided, with the stability that improves enzyme, make things convenient for goods to transport, make the zymin that makes quickly dissolving, and dissolving back enzyme liquid is limpid, the clarity height in water.
For solveing the technical problem, the technical solution used in the present invention is, a kind of production method of high enzymic activity instant enzyme preparation, its content is: in the last handling process to fermented liquid behind high density fermentation, the fermented liquid that separates after concentrating is carried out albumen salt precipitation and vacuum lyophilization.
The production method of above-mentioned high enzymic activity instant enzyme preparation, it comprises the following steps:
(1) after the fermentation ends, fermented liquid is moved to storage tank and separates with membrane filtration and handle;
(2) in above-mentioned treatment solution, add food grade ammonium sulfate while stirring, leave standstill after the stirring;
(3) separate above-mentioned treatment solution, collecting precipitation with whizzer; The gained precipitation is filled in the charging tray of vacuum freeze drier;
(4) start vacuum freeze drier and carry out vacuum lyophilization; Through pulverize, weigh and pack finished product.
The production method of above-mentioned high enzymic activity instant enzyme preparation, its particular content is:
(1) after the fermentation ends, fermented liquid is moved to storage tank; Yeast or ferment product carry out membrane microfiltration, concentrated, the membrane microfiltration processing of membrane ultrafiltration again through after the centrifugation; Membrane ultrafiltration concentrates mold fermentation liquid, membrane microfiltration through carrying out behind the filter press again;
(2) add food grade ammonium sulfate while stirring in above-mentioned treatment solution, addition is 22%～47% of a feed liquid quality, stirs to leave standstill 3.5～7 hours after 30～60 minutes;
(3) separate above-mentioned treatment solution, collecting precipitation with interior with whizzer at inlet amount 4L/min; The gained precipitation is filled in the charging tray of vacuum freeze drier;
(4) directly open vacuum pump behind the startup vacuum freeze drier and carry out vacuum lyophilization, after 20～40 minutes material temperature drops are to-30 ℃, start the shelf heating, the thermal oil temperature is made as 60 ℃, heats 7～10 hours to expecting that temperature stops vacuum lyophilization when reaching 25 ℃～30 ℃; Through pulverize, weigh and pack finished product.
The described whizzer of step (3) is placed in-line 2 tubular-bowl centrifuges; The charging thickness of described charging tray is 1 centimetre.
Method of the present invention is used for the production zymin, can improve enzyme stability, make things convenient for goods to transport.The enzyme activity height of gained enzyme preparation product, be fermentation broth enzyme live more than 25 times, product active ingredient is lost seldom, can keep the original physics of material, chemical property, water content in 3%, the long preservative period of goods, good stability; Quickly dissolving in water, and dissolving back enzyme liquid is limpid, the clarity height, and use more convenient.
Embodiment
The present invention is described in more detail below in conjunction with embodiment.
Embodiment 1
A kind of production method of high enzymic activity instant enzyme preparation comprises:
(1) after the fermentation ends, ferment product is moved to storage tank.Open disc separator, after machine to be separated reaches at full speed, open feed valve, and according to the difference of fermented liquid concentration, regulate inlet amount or change the nozzle in different apertures.
(2) above-mentioned separating machine clear liquid is carried out micro-filtration, ultrafiltration with membrane filtration system, ultrafiltrated carries out micro-filtration once more.
(3) add food grade ammonium sulfate while stirring in above-mentioned treatment solution, addition is 47% of a feed liquid quality, stirs to leave standstill 3.5 hours after 30 minutes.
(4) separate above-mentioned treatment solution, collecting precipitation at inlet amount 4L/min with 2 tubular-bowl centrifuges that interior usefulness is together in series.
(5) above-mentioned precipitation is filled in the charging tray of vacuum freeze drier, charging thickness is 1 centimetre.
(6) start vacuum freeze drier, the thermal oil temperature is made as 60 ℃, opens vacuum pump.
Start the shelf heating after (7) 30 minutes, heat 8 hours to expecting that temperature stops vacuum lyophilization when reaching 25 ℃ ~ 30 ℃.
(8) through pulverizing, weigh and packing, be finished product.
Embodiment 2
A kind of production method of high enzymic activity instant enzyme preparation comprises:
(1) after the fermentation ends, mold fermentation liquid is moved to storage tank.Open sheet frame oil-pressure pump power supply, make each parts of sheet frame become impaction state, keep sheet frame pressure 12 ~ 15MPa to stop power supply.Open sheet frame feed valve door, open each clear liquid discharging valve of sheet frame.When material enters sheet frame more and more slowly the time, when feed pressure reaches 4MPa, stop charging.
(2) the above-mentioned press filtration stillness of night is carried out ultrafiltration with membrane filtration system, ultrafiltrated carries out micro-filtration once more.
(3) add food grade ammonium sulfate while stirring in above-mentioned treatment solution, addition is 22% of a feed liquid quality, stirs to leave standstill 7 hours after 1 hour.
(4) separate above-mentioned treatment solution, collecting precipitation at inlet amount 4L/min with 2 tubular-bowl centrifuges that interior usefulness is together in series.
(5) above-mentioned precipitation is filled in the charging tray of vacuum freeze drier, charging thickness is 1 centimetre.
(6) start vacuum freeze drier, the thermal oil temperature is made as 60 ℃, opens vacuum pump.
The material temperature drop starts the shelf heating to-30 ℃ after (7) 20 minutes, heats 10 hours to expecting that temperature stops vacuum lyophilization when reaching 25 ℃ ~ 30 ℃.
(8) through pulverizing, weigh and packing, be finished product.
Embodiment 3
A kind of production method of high enzymic activity instant enzyme preparation comprises:
(1) after the fermentation ends, yeast fermentation broth is moved to storage tank.Open disc separator, machine to be separated can be opened feed valve after reaching at full speed, according to the difference of fermented liquid concentration, regulates inlet amount or changes the nozzle in different apertures.
(2) the above-mentioned separating machine stillness of night is carried out micro-filtration, ultrafiltration with membrane filtration system, ultrafiltrated carries out micro-filtration once more.
(3) add food grade ammonium sulfate while stirring in above-mentioned treatment solution, addition is 35% of a feed liquid quality, stirs to leave standstill 6 hours after 50 minutes.
(4) separate above-mentioned treatment solution, collecting precipitation at inlet amount 4L/min with 2 tubular-bowl centrifuges that interior usefulness is together in series.
(5) above-mentioned precipitation is filled in the charging tray of vacuum freeze drier, charging thickness is 1 centimetre.
(6) start vacuum freeze drier, the thermal oil temperature is made as 60 ℃, opens vacuum pump.
The material temperature drop starts the shelf heating to-30 ℃ after (7) 40 minutes, heats 7 hours to expecting that temperature stops vacuum lyophilization when reaching 25 ℃ ~ 30 ℃.
(8) through pulverizing, weigh and packing, be finished product.
Embodiment 4
The check of gained finished product in the various embodiments described above:
A, Oranoleptic indicator:
Color and luster: be light green or slightly lurid powder.
Smell: do not have bad smell.
B, physical and chemical index:
Water content (%)≤3;
Enzyme work 〉=fermenting enzyme running water flat * 25;
Solvability: speed is water-soluble, and dissolving back solution is limpid transparent.
C, sanitary index:
Arsenic (in As)≤0.1mg/kg;
Plumbous (in Pb)≤0.2mg/kg;
Cadmium (in Cd)≤0.1mg/kg;
Aflatoxin B1≤5ug/kg;
Coliform≤500/100g;
Salmonellas: do not detect.
Claims (4)
1. the production method of a high enzymic activity instant enzyme preparation, its content is: in the last handling process to fermented liquid behind high density fermentation, the fermented liquid that separates after concentrating is carried out albumen salt precipitation and vacuum lyophilization.
2. production method according to claim 1, it comprises the following steps:
(1) after the fermentation ends, fermented liquid is moved to storage tank and separates with membrane filtration and handle;
(2) in above-mentioned treatment solution, add food grade ammonium sulfate while stirring, leave standstill after the stirring;
(3) separate above-mentioned treatment solution, collecting precipitation with whizzer; The gained precipitation is filled in the charging tray of vacuum freeze drier;
(4) start vacuum freeze drier and carry out vacuum lyophilization; Through pulverize, weigh and pack finished product.
3. production method according to claim 2, its particular content is:
(1) after the fermentation ends, fermented liquid is moved to storage tank; Yeast or ferment product carry out membrane microfiltration, concentrated, the membrane microfiltration processing of membrane ultrafiltration again through after the centrifugation; Membrane ultrafiltration concentrates mold fermentation liquid, membrane microfiltration through carrying out behind the filter press again;
(2) add food grade ammonium sulfate while stirring in above-mentioned treatment solution, addition is 22%～47% of a feed liquid quality, stirs to leave standstill 3.5～7 hours after 30～60 minutes;
(3) separate above-mentioned treatment solution, collecting precipitation with interior with whizzer at inlet amount 4L/min; The gained precipitation is filled in the charging tray of vacuum freeze drier;
(4) directly open vacuum pump behind the startup vacuum freeze drier and carry out vacuum lyophilization, after 20～40 minutes material temperature drops are to-30 ℃, start the shelf heating, the thermal oil temperature is made as 60 ℃, heats 7～10 hours to expecting that temperature stops vacuum lyophilization when reaching 25 ℃～30 ℃; Through pulverize, weigh and pack finished product.
4. production method according to claim 3, the described whizzer of its step (3) are placed in-line 2 tubular-bowl centrifuges; The charging thickness of described charging tray is 1 centimetre.
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| CN201010188024A CN101870965A (en) | 2010-06-01 | 2010-06-01 | Production method of high enzymic activity instant enzyme preparation |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN85103913A (en) * | 1985-06-19 | 1986-12-24 | 国家医药管理局上海医药工业研究院 | Technology for making collagenase |
| CN1724659A (en) * | 2005-06-14 | 2006-01-25 | 浙江大学 | Preparation method of chitin incision enzyme |
-
2010
- 2010-06-01 CN CN201010188024A patent/CN101870965A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN85103913A (en) * | 1985-06-19 | 1986-12-24 | 国家医药管理局上海医药工业研究院 | Technology for making collagenase |
| CN1724659A (en) * | 2005-06-14 | 2006-01-25 | 浙江大学 | Preparation method of chitin incision enzyme |
Non-Patent Citations (2)
| Title |
|---|
| 刘军.: "膜分离技术在生物化工中的应用", 《科技信息（学术研究）》, no. 9, 31 December 2007 (2007-12-31), pages 84 - 85 * |
| 董充慧等.: "真空冷冻干燥技术在生物制药方面的应用", 《沈阳药科大学学报》, vol. 26, 31 July 2009 (2009-07-31), pages 76 - 78 * |
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Address after: Miao road Laoshan District 266061 Shandong city of Qingdao province Shandong No. 29 high building 12A07 Applicant after: Qingdao Weilan Biology Group Co., Ltd. Address before: Miao road Laoshan District 266061 Shandong city of Qingdao Province, No. 29 building 6 floor, room 607, Shandong high speed Applicant before: Qingdao Continent Biotech Co., Ltd. |
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Application publication date: 20101027 |