CN105349419A - Granular immobilized enzyme preparation equipment and application method - Google Patents

Granular immobilized enzyme preparation equipment and application method Download PDF

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Publication number
CN105349419A
CN105349419A CN201510979229.9A CN201510979229A CN105349419A CN 105349419 A CN105349419 A CN 105349419A CN 201510979229 A CN201510979229 A CN 201510979229A CN 105349419 A CN105349419 A CN 105349419A
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China
Prior art keywords
immobilized enzyme
preparation equipment
particulate state
state immobilized
enzyme according
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CN201510979229.9A
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Chinese (zh)
Inventor
马士忠
胡永红
李莉
马飞鸿
杜向龙
胡伟
马士强
马道功
杨召鹏
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SHANGHAI HANHONG CHEMICAL INDUSTRY Co Ltd
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SHANGHAI HANHONG CHEMICAL INDUSTRY Co Ltd
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Priority to CN201510979229.9A priority Critical patent/CN105349419A/en
Publication of CN105349419A publication Critical patent/CN105349419A/en
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/12Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M27/00Means for mixing, agitating or circulating fluids in the vessel
    • C12M27/02Stirrer or mobile mixing elements
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M29/00Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/06Nozzles; Sprayers; Spargers; Diffusers

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  • Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Sustainable Development (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Thermal Sciences (AREA)
  • Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

The invention relates to granular immobilized enzyme preparation equipment and an application method. The granular immobilized enzyme preparation equipment comprises a reducer, a stirring device, a heating jacket, a barrel body and a spray nozzle, wherein the reducer is connected above the barrel body; the stirring device is arranged in the barrel body; the heating jacket is arranged outside the barrel body; the spray nozzle is connected to the lower part of the barrel body and is a conical spray nozzle. The granular immobilized enzyme preparation equipment and the application method have the benefits as follows: the device can realize mixing and heating mixed functions, materials such as carrageenan, gelatin and the like are separately blended with immobilized enzyme or immobilized bacteria, and by the aid of heating stirring and cooling, the granular immobilized enzyme or immobilized bacteria can be obtained.

Description

The Preparation equipment of particulate state immobilized enzyme and using method
Technical field
The present invention relates to biochemical field, particularly relate to the reactor producing immobilized enzyme, specifically refer to a kind of Preparation equipment and using method of particulate state immobilized enzyme.
Background technology
Modern biotechnology achieves the scale operation of enzyme, and free enzyme is widely applied in food, chemical industry and medicine industry.Although enzyme can catalyzed reaction efficiently under normal temperature and pressure conditions, and has very high specificity, enzyme as a kind of protein, poor stability, all unstable to heat, strong acid, highly basic and organic solvent etc. [36]even if, under the optimum reaction conditions of enzyme, also often very fast inactivation.And free enzyme can only adopt batch reactor, after reaction, enzyme can not reclaim, and makes product separation purification difficult, easily causes protein contamination, so be not also a kind of desirable catalyzer concerning free enzyme modern industry.For this reason, people have prepared all kinds of immobilized enzyme.In all kinds of immobilization material, carrageenin, gelatin have good stability, advantage that cost is low, are consumption immobilization materials the most widely.But the glue after immobilization is generally large bulk, need manual or machinery section, and sheet catalyst loads and unloads into reactor inconvenience, laminar flow is poor, easy fluid short circuit etc. in reactor, and section too increases operation and causes the loss of catalyzer.
Spherical catalyst has advantage widely, can carrageeenen immobilized enzyme be changed into spherical by general sheet? have researchist to do some to attempt.Illustrate based on water incompatible with many organic solvents if any document, water is dispersed into the principle of drop in organic solvent, (Sun Wanru, microbiology is circulated a notice of, 1986,13 (1): 213-35) report uses butylacetate dispersion carrageenin liquid, forms spheroidal particle.Concrete grammar is as follows: the carrageenan solutions adding hot preparation 8%, 60 DEG C are cooled under room temperature, the enchylema getting 20ml glue and 20ml40% mixes, stream is added in the butylacetate of insulation 30 DEG C of stirring, until be dispersed into small-particle, add the KCl solution of 10%, continue stirring 10 minutes elimination solvents, then the KCl adding 2% solidifies the immobilized cell that namely a hour form 1 ~ 3mm.The shortcoming of this method is that the butylacetate used not easily reclaims, and solvent has people irritant, needs to operate under the environment that security is higher.
But also there is no at present industrial can cheaply on a large scale preparation with the Apparatus and method for of gelatin, particulate state immobilized enzyme that carrageenin is material or immobilized cell generally.
Summary of the invention
The object of the invention is the shortcoming overcoming above-mentioned prior art, provide a kind of Preparation equipment and the using method that can prepare particulate state immobilized enzyme on a large scale.
To achieve these goals, the present invention has following formation:
The Preparation equipment of this particulate state immobilized enzyme, its principal feature is, the Preparation equipment of described particulate state immobilized enzyme comprises step-down gear, whipping appts, heating jacket, cylindrical shell and shower nozzle, described step-down gear is connected with above described cylindrical shell, whipping appts is provided with in described cylindrical shell, be provided with heating jacket outside described cylindrical shell, be connected with shower nozzle below described cylindrical shell, wherein said shower nozzle is Conic nozzle.
Preferably, described whipping appts is paddle paddle.
More preferably, described heating jacket is built-in with heating medium, and described heating medium is water and steam, and Heating temperature is 20 ~ 120 DEG C.
Further preferably, described Conic nozzle is distributed with circular hole or the aciculiform shower nozzle of 0.5 ~ 2mm, and described Conic nozzle aperture is 0.8 ~ 1.8mm, and pitch of holes is 6 ~ 15mm.
Still more preferably, described Conic nozzle aperture is 1.0 ~ 1.4mm, and pitch of holes is 8 ~ 12mm.
Again further preferably, described whipping appts rotating speed is 150 ~ 350rpm.
Most preferably, described whipping appts rotating speed is 200 ~ 300rpm.
According to the using method of the Preparation equipment of described particulate state immobilized enzyme, its principal feature is, the Preparation equipment of the particulate state immobilized enzyme described in described using method uses prepares particulate state immobilized enzyme or immobilized cell.
Preferably, described using method is specially:
By immobilization material and thalline or enzyme organism, dissolve batching in advance, then the Preparation equipment of described particulate state immobilized enzyme is metered into through pump, through Hybrid Heating, enter shower nozzle, under the rotation of shower nozzle and under action of gravity, dispersion becomes spherical droplets, and falling into below solidified liquid becomes spherical solid particles.
More preferably, described immobilization material comprises: gelatin, carrageenin, sodium alginate, polyvinyl alcohol, agar, and the mixture of mixing.
Have employed Preparation equipment and the using method of the particulate state immobilized enzyme in this invention, its beneficial effect is: this device can reach mixing and the mixing functions heated, carrageenin, the materials such as gelatin and immobilized enzyme or immobilized thallus are prepared burden separately, carrageenin, gelatin zero pour is all below 50 degree, higher than being liquid state on zero pour, mix with biomaterial (enzyme or cell), through being uniformly mixed and being incubated, flow to Conic nozzle, under the Conic nozzle effect rotated, dispersion becomes spherical liquid, the liquid of ejection starts to be liquid stream, then because surface tension, become drop gradually to fall, fall into below solidified liquid to solidify, cooled and solidified becomes solid, particulate state immobilized enzyme or immobilized thallus can be obtained.
Accompanying drawing explanation
Fig. 1 is the Preparation equipment schematic diagram of particulate state immobilized enzyme of the present invention.
Fig. 2 is the use schematic diagram of Preparation equipment for the preparation of immobilized enzyme or immobilized cell of particulate state immobilized enzyme of the present invention.
Reference numeral
1 step-down gear
2 paddles
3 heating jackets
4 cylindrical shells
5 shower nozzles
A immobilization material batching reactor
B biomaterial batching reactor
C particulate state immobilized enzyme Preparation equipment
D solid solution hold-up vessel
Embodiment
In order to more clearly describe technology contents of the present invention, conduct further description below in conjunction with specific embodiment.
The Preparation equipment of the particulate state immobilized enzyme as shown in Fig. 1 ~ 2 and using method, its specific embodiment is:
Embodiment 1:
Take 2.0kgK-carrageenin and 50L water adds in A reactor, heating in water bath to 90 DEG C, glue is melted, is incubated for subsequent use; Take L-Aminoacylase 0.5KG and water 50L adds B reactor, 50 DEG C of heat preservation for standby use.Reactor rotating speed is 230rpm, open pore size is 1.2mm, start the C particulate state immobilized enzyme Preparation equipment of cumulative volume 50L, below discharge valve temporary close, A liquid and B liquid add C particulate state immobilized enzyme Preparation equipment according to 2:1 ratio, holding temperature sets 80 DEG C, stir 10 minutes, Open valve, control rotating speed is 230rpm, in the D solid solution hold-up vessel of the drop instillation below 0.3mol/LKCI solution of ejection, after 8 hours, filtering, obtain the spheroidal particle of diameter about 1mm, being greater than 95% through measuring 1mm particle total amount.
Embodiment 2:
Take 5.0kg gelatin and 100L water adds in A reactor, 120 DEG C, glue sterilizing is melted, be cooled to 70 DEG C of insulations for subsequent use; Take L-Aminoacylase 0.5KG and water 30L adds B reactor, 50 DEG C of heat preservation for standby use.Reactor rotating speed is 230rpm, and open pore size is 1.2mm, starts the C particulate state immobilized enzyme Preparation equipment of cumulative volume 50L, below discharge valve temporary close, A liquid and B liquid add C particulate state immobilized enzyme Preparation equipment according to 3:1 ratio, and holding temperature sets 80 DEG C, stir 10 minutes, Open valve, control rotating speed is 230rpm, is chilled in 5 DEG C of water in advance under the drop instillation of ejection, after 8 hours, filtering, obtain the spheroidal particle of diameter about 1mm, being greater than 85% through measuring 1mm particle total amount.
Embodiment 3:
Take agar 5KG and 100L water adds in A reactor, 120 DEG C, glue sterilizing is melted, be cooled to 50 DEG C of insulations for subsequent use; Take L-Aminoacylase 0.5KG and water 30L adds B reactor, 50 DEG C of heat preservation for standby use.Reactor rotating speed is 230rpm, and open pore size is 1.2mm, starts the C particulate state immobilized enzyme Preparation equipment of cumulative volume 50L, below discharge valve temporary close, A liquid and B liquid add C particulate state immobilized enzyme Preparation equipment according to 3:1 ratio, and holding temperature sets 80 DEG C, stir 10 minutes, Open valve, control rotating speed is 230rpm, is chilled in 5 DEG C of water in advance under the drop instillation of ejection, after 8 hours, filtering, obtain the spheroidal particle of diameter about 1mm, being greater than 91% through measuring 1mm particle total amount.
Embodiment 4:
Weighing polyvinyl alcohol 8KG and 90L water add in A reactor, 120 DEG C, and glue sterilizing is melted, and are cooled to 30 DEG C of insulations for subsequent use; Take L-Aminoacylase 0.5KG and water 30L adds B reactor, 30 DEG C of heat preservation for standby use.Reactor rotating speed is 230rpm, open pore size is 1.2mm, start the C particulate state immobilized enzyme Preparation equipment of cumulative volume 50L, below discharge valve temporary close, A liquid and B liquid add C particulate state immobilized enzyme Preparation equipment according to 3:1 ratio, holding temperature sets 80 DEG C, stir 10 minutes, Open valve, control rotating speed is 230rpm, the drop instillation of ejection is containing 25g/L borax, in the solution D solid solution hold-up vessel of 50g/L sodium-chlor, after 3 hours, filter, obtaining the spheroidal particle of diameter about 1mm, being greater than 90% through measuring 1mm particle total amount.
Embodiment 5:
Take 2.0kgK-carrageenin and 50L water adds in A reactor, heating in water bath, to 9O DEG C, makes glue melt, and is incubated for subsequent use; Take L-Aminoacylase 0.5KG and water 50L adds B reactor, 50 DEG C of heat preservation for standby use.Reactor rotating speed is 230rpm, open pore size is 3mm, start the C particulate state immobilized enzyme Preparation equipment of cumulative volume 50L, below discharge valve temporary close, A liquid and B liquid add C particulate state immobilized enzyme Preparation equipment according to 2:1 ratio, holding temperature sets 80 DEG C, stir 10 minutes, Open valve, control rotating speed is 230rpm, in the D solid solution hold-up vessel of the drop instillation below 0.3mol/LKCl solution of ejection, after 8 hours, filter, obtain the spheroidal particle of diameter about 1mm ~ 4mm, 25% is less than through measuring 1mm particle total amount, more than 3mm particle is greater than 50%, there is particle adhesion phenomenon.
Embodiment 6:
Weighing polyvinyl alcohol 8KG and 90L water add in A reactor, 120 DEG C, and glue sterilizing is melted, and are cooled to 30 DEG C of insulations for subsequent use; Take L-Aminoacylase 0.5KG and water 30L adds B reactor, 30 DEG C of heat preservation for standby use.Reactor rotating speed is 230rpm, open pore size is 0.6mm, start the C particulate state immobilized enzyme Preparation equipment of cumulative volume 50L, below discharge valve temporary close, A liquid and B liquid add C particulate state immobilized enzyme Preparation equipment according to 3:1 ratio, holding temperature sets 80 DEG C, stir 10 minutes, Open valve, control rotating speed is 230rpm, the drop instillation of ejection is containing 25g/L borax, in the solution D solid solution hold-up vessel of 50g/L sodium-chlor, after 3 hours, filter, obtain the spheroidal particle of diameter about 0.4mm ~ 1mm, be less than 0.4mm particle total amount through mensuration diameter and be greater than 30%.
Embodiment 7:
Take sodium alginate 3KG and 100L water adds in A reactor, 120 DEG C, sterilizing is melted, be cooled to 50 DEG C of insulations for subsequent use; Take L-Aminoacylase 0.5KG and water 30L adds B reactor, 50 DEG C of heat preservation for standby use.Reactor rotating speed is 230rpm, open pore size is 1.2mm, start the C particulate state immobilized enzyme Preparation equipment of cumulative volume 50L, below discharge valve temporary close, A liquid and B liquid add C particulate state immobilized enzyme Preparation equipment according to 3:1 ratio, and holding temperature sets 50 DEG C, stir 10 minutes, Open valve, control rotating speed is 230rpm, is chilled to the 2%CaCl of 5 DEG C under the drop instillation of ejection in advance 2in the D solid solution hold-up vessel of solution, after 2 hours, filtering, obtain the spheroidal particle of diameter about 1mm, being greater than 96% through measuring 1mm particle total amount.
Embodiment 8:
Take sodium alginate 3KG and 100L water adds in A reactor, 120 DEG C, sterilizing is melted, be cooled to 50 DEG C of insulations for subsequent use; Take L-Aminoacylase 0.5KG and water 30L adds B reactor, 50 DEG C of heat preservation for standby use.Reactor rotating speed is 100rpm, open pore size is 1.2mm, start the C particulate state immobilized enzyme Preparation equipment of cumulative volume 50L, below discharge valve temporary close, A liquid and B liquid add C particulate state immobilized enzyme Preparation equipment according to 3:1 ratio, and holding temperature sets 50 DEG C, stir 10 minutes, Open valve, control rotating speed is 230rpm, is chilled to the 2%CaCl of 5 DEG C under the drop instillation of ejection in advance 2in the D solid solution hold-up vessel of solution, after 2 hours, filter, obtain the spheroidal particle of diameter about 1 ~ 3mm, be less than 30%, 2mm particle total amount through mensuration 1mm particle total amount and be greater than 30%, have particle bond situation.
Embodiment 9:
Take sodium alginate 3KG and 100L water adds in A reactor, 120 DEG C, sterilizing is melted, be cooled to 50 DEG C of insulations for subsequent use; Take L-Aminoacylase 0.5KG and water 30L adds B reactor, 50 DEG C of heat preservation for standby use.Reactor rotating speed is 400rpm, open pore size is 1.2mm, start the C particulate state immobilized enzyme Preparation equipment of cumulative volume 50L, below discharge valve temporary close, A liquid and B liquid add C particulate state immobilized enzyme Preparation equipment according to 3:1 ratio, and holding temperature sets 50 DEG C, stir 10 minutes, Open valve, control rotating speed is 230rpm, is chilled to the 2%CaCl of 5 DEG C under the drop instillation of ejection in advance 2in the D solid solution hold-up vessel of solution, after 2 hours, filter, obtain diameter 0.3 ~ 1mm spheroidal particle, be greater than 34% through measuring 0.3mm particle total amount, be greater than 0.8mm particle total amount and be less than 30%, particle is less than normal.
Have employed Preparation equipment and the using method of the particulate state immobilized enzyme in this invention, its beneficial effect is: this device can reach mixing and the mixing functions heated, carrageenin, the materials such as gelatin and immobilized enzyme or immobilized thallus are prepared burden separately, carrageenin, gelatin zero pour is all below 50 degree, higher than being liquid state on zero pour, mix with biomaterial (enzyme or cell), through being uniformly mixed and being incubated, flow to Conic nozzle, under the Conic nozzle effect rotated, dispersion becomes spherical liquid, the liquid of ejection starts to be liquid stream, then because surface tension, become drop gradually to fall, fall into below solidified liquid to solidify, cooled and solidified becomes solid, particulate state immobilized enzyme or immobilized thallus can be obtained.
In this description, the present invention is described with reference to its specific embodiment.But, still can make various amendment and conversion obviously and not deviate from the spirit and scope of the present invention.Therefore, specification sheets and accompanying drawing are regarded in an illustrative, rather than a restrictive.

Claims (10)

1. the Preparation equipment of a particulate state immobilized enzyme, it is characterized in that, the Preparation equipment of described particulate state immobilized enzyme comprises step-down gear, whipping appts, heating jacket, cylindrical shell and shower nozzle, described step-down gear is connected with above described cylindrical shell, whipping appts is provided with in described cylindrical shell, be provided with heating jacket outside described cylindrical shell, be connected with shower nozzle below described cylindrical shell, wherein said shower nozzle is Conic nozzle.
2. the Preparation equipment of particulate state immobilized enzyme according to claim 1, is characterized in that, described whipping appts is paddle paddle.
3. the Preparation equipment of particulate state immobilized enzyme according to claim 1, is characterized in that, described heating jacket is built-in with heating medium, and described heating medium is water and steam, and Heating temperature is 20 ~ 120 DEG C.
4. the Preparation equipment of particulate state immobilized enzyme according to claim 1, is characterized in that, described Conic nozzle is distributed with circular hole or the aciculiform shower nozzle of 0.5 ~ 2mm, and the aperture of described Conic nozzle is 0.8 ~ 1.8mm, and pitch of holes is 6 ~ 15mm.
5. the Preparation equipment of particulate state immobilized enzyme according to claim 4, is characterized in that, the aperture of described Conic nozzle is 1.0 ~ 1.4mm, and pitch of holes is 8 ~ 12mm.
6. the Preparation equipment of particulate state immobilized enzyme according to claim 1, is characterized in that, the rotating speed of described whipping appts is 150 ~ 350rpm.
7. the Preparation equipment of particulate state immobilized enzyme according to claim 6, is characterized in that, the rotating speed of described whipping appts is 200 ~ 300rpm.
8. the using method of the Preparation equipment of the particulate state immobilized enzyme according to any one of claim 1 ~ 7, it is characterized in that, the Preparation equipment of the particulate state immobilized enzyme described in described using method uses prepares particulate state immobilized enzyme or immobilized cell.
9. the using method of the Preparation equipment of particulate state immobilized enzyme according to claim 8, is characterized in that, described using method is specially:
By immobilization material and thalline or enzyme organism, dissolve batching in advance, then the Preparation equipment of described particulate state immobilized enzyme is metered into through pump, through Hybrid Heating, enter shower nozzle, under the rotation of shower nozzle and under action of gravity, dispersion becomes spherical droplets, and falling into below solidified liquid becomes spherical solid particles.
10. the using method of the Preparation equipment of particulate state immobilized enzyme according to claim 9, is characterized in that, described immobilization material comprises: gelatin, carrageenin, sodium alginate, polyvinyl alcohol, agar, and the mixture of mixing.
CN201510979229.9A 2015-12-22 2015-12-22 Granular immobilized enzyme preparation equipment and application method Pending CN105349419A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113122445A (en) * 2021-05-06 2021-07-16 南京普瑞特生物科技有限公司 Equipment for preparing optically pure 1- (1-naphthyl) ethylamine by splitting with immobilized enzyme method and use method

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113122445A (en) * 2021-05-06 2021-07-16 南京普瑞特生物科技有限公司 Equipment for preparing optically pure 1- (1-naphthyl) ethylamine by splitting with immobilized enzyme method and use method

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