CN104531804A - Method for production of crystal trehalose and co-production of moisturizing syrup - Google Patents
Method for production of crystal trehalose and co-production of moisturizing syrup Download PDFInfo
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Abstract
The invention relates to a method for production of crystal trehalose and co-production of moisturizing syrup. Trehalose synthase immobilization is carried out by adopting a nanofiber; high temperature and high sugar concentration are adopted to carry out enzymic preparation reaction; maltose and isomaltose of affecting trehalose crystalline purity are removed by adopting a two-enzyme method; the purity of a trehalose crystal product is improved; the moisturizing syrup is obtained when the high-purity trehalose is prepared by a chromatographic ternary component separation technology; efficient preparation of trehalose and comprehensive utilization of a separating medium are achieved in the overall process; the immobilized trehalose synthase can be used for a plurality of times; the use frequency exceeds 100 times; the use efficiency of enzyme is greatly improved; the maltose and isomaltose of affecting trehalose crystallization are removed; the product with the purity being greater than 99% can be obtained by crystallization once; the moisturizing syrup is produced when the trehalose is produced; production free of a side product is achieved; the by-product is produced by the method instead of the traditional extraction mode; the comprehensive utilization value is improved; separation of three components is achieved by the ternary component separation technology; the traditional twice component separation mode is replaced; and the method is convenient to operate, and has a wide industrialized prospect.
Description
Technical field
The present invention relates to a kind of production method of crystalline trehalose coproduction moisture-keeping syrup, belong to bioengineering field.
Background technology
Trehalose (Trehalose) passes through α by two glucose molecules, α-1, the non-reducing disaccharide that 1 glycosidic link connects, this product is a kind of safe and reliable natural carbohydrate, trehalose all extensively exists in the many edible animals and plants of occurring in nature and microbe, as having trehalose in mushroom class edible in people's daily life, seaweeds, beans, shrimp, bread, beer and yeast fermentation food.Trehalose is generally acknowledged safety food (GRAS), calendar year 2001, united States food and drug administration (FDA) authorizes trehalose as generally regarded as safe food, shows direct-edible or as food ingredients in " trehalose " (GB/T23529-2009) national standard.
Trehalose is widely used in the aspects such as food-processing, agricultural, makeup, healthcare products and field of medicaments.
In the food industry, because of the characteristic such as sweet taste and energy supply of the irreducibility of trehalose, humidity-preserving type, anti-freeze and drying property, high-quality, trehalose is used as foodstuff additive or sweetening agent, some desiccating food products can be made after regaining moisture still to keep original shape, color and quality are the same with fresh provisions.
In medicines and health protection field, trehalose can be used as the stablizer of reagent medicine and diagnosis medicine, can be used as the conserving liquid of transplant organ, as stablizer or the quality improver of the various biological products such as lymph, hormone, vaccine, microbiotic, VITAMIN, enzyme.Oral trehalose can also improve osteoporosis.
In biology field, the biologically active substances such as trehalose dry antibody, thrombocyte, enzyme, virus, without the need to freezen protective, all can rejuvenate after rehydration.Trehalose replaces blood plasma as the stablizer of biological products, can not only preserve by normal temperature, also can prevent the hepatitis B that causes because of blood contamination and aids transmission, thus ensure quality and the safety of biological products.
In cosmetic industry, utilize the performance of keeping humidity of trehalose, the sulfate derivative of trehalose for skin cosmetics, as skin detergents, skin moisturizer, uv-absorbing agent, the quality improver, stablizer etc. of lipstick.
At agriculture field, what built by biotechnology contains trehalose transgenic plant, opens new way for cultivating drought resisting freeze resistant transgenic plant.Crop containing trehalose is not only drought-enduring resistance toly to be frozen, and seems fresher after results processing, has more local flavor.
At present, world market being in great demand to trehalose, estimates that the annual requirement in a few years from now on will reach more than 150,000 tons, along with the quick expansion of Application Areas, to the demand of trehalose also by corresponding increase.
The method mainly microorganism extraction process of the production trehalose of researching and developing about trehalose both at home and abroad, microbe fermentation method, enzymatic clarification trehalose etc.Wherein, microorganism extraction process, and extract from bread yeast, cereuisiae fermentum, be the main method of producing trehalose at present.Double-enzyme method trehalose synthesis, comprises multistep enzyme digestion reaction, and namely utilize multiple enzyme handle, maltose or glucose sugar to be converted into trehalose, reaction process temperature is low, concentration is low.(1) double-enzyme method: the synergy utilizing malt oligosaccharide based mycose synthetase and malt oligosaccharide based mycose lytic enzyme, the straight chain of certain chain length is converted into trehalose, join 20%(w/w) maltose solution → enzyme-added (2U/g maltose), 15 DEG C, pH7.0, reaction 48h → enzyme → filtration → decolouring of going out → from friendship → concentrated → crystallization → washing → finished product; (2) extraction method: cell → ultra micro membrane filtration → condensing crystal that slant strains → seed culture → fermentation culture → collection thalline → dehydration → drying → pulverizing → dissolving is pulverized; (3) fermentation method: slant strains → seed culture → fermentation culture → filtration sterilization → decolouring → from friendship → concentrated → crystallization → washing → finished product.
These above-mentioned methods are more loaded down with trivial details, have that trehalose preparation efficiency is low, reaction density is low, the temperature of reaction is low, produce the shortcomings such as the enzyme of trehalose or thalline can not recycle, complicated operation.
Summary of the invention
The present invention is directed to prior art deficiency, a kind of production method of crystalline trehalose coproduction moisture-keeping syrup is provided.
This technology is passed through trehalose synthase immobilization, utilize nanofiber to being fixed of TreP, take high temperature, high dense conversion high maltose syrup generation trehalose, control from many-sides such as temperature, PH, concentration, finally obtain higher trehalose output, can continuous prodution trehalose efficiently, there is wide industrial prospect.
Two enzyme united hydrolysis method is adopted in this technology treating process, the maltose and isomaltose that affect trehalose crystallization are eliminated, due in early stage saccharifying, maltose in the thick liquid of trehalose is suitable with the molecular weight of isomaltose and trehalose, adopt the treatment process of physics not remove, there is considerable influence to secondary crystallization, the two is deposited and easily causes the purity of primary crystallization inadequate 99% in case, sometimes need to carry out secondary recrystallization, just can bring up to more than 99%.Follow-up employing three components chromatographic separation technology, improves the purity of trehalose further and reclaims glucose and polysaccharide, produces a kind of moisture-keeping syrup of trehalose product coproduction simultaneously and glucose.
Technical problem to be solved by this invention mainly solves the problem that trehalose synthase can not recycle, novel method makes zymin can recycle more than 100 times, achieves the once preparation of zymin, repeatedly recycles, reduce the energy consumption of producing, improve the enzyme service efficiency of trehalose; Solve the present situation that in enzymatic conversion process, temperature of reaction is low, reaction density is low simultaneously, improve the utilising efficiency of equipment; Solve the problem that in existing trehalose crystallization, product purity is low simultaneously, realize primary crystallization and can obtain the product that purity is greater than 99%, higher than the standard of existing product purity 98%, improving product purity.
Technical scheme of the present invention is as follows:
A production method for crystalline trehalose coproduction moisture-keeping syrup, comprises the steps:
(1) epoxidation composite nano fiber is adopted to process being fixed of TreP;
(2) be substrate with high maltose syrup, utilize immobilization TreP to produce trehalose, when content of trehalose reaches 55%, terminate this reaction;
(3) adopt double-enzyme composite hydrolysis method of purification, the trehalose obtain step (2) and the mixing solutions of maltose carry out purification process, remove the maltose and the isomaltose that affect trehalose crystallization purity, improve the purity of trehalose crystalline product;
(4) sugar component more than trehalose, glucose and trisaccharide in the mixing solutions adopting chromatogram ternary component isolation technique division step (3) to obtain, object improves the purity of trehalose, reclaim the sugar component of more than glucose and trisaccharide, a kind of moisture-keeping syrup of coproduction while production trehalose product.
Preferred according to the present invention, in described step (1), adopt the step of epoxidation composite nano fiber to being fixed of TreP as follows: to get enzyme and live as the TreP enzyme liquid 90-110ml of 800-1500U/ml, add after being diluted to 290-310ml with the phosphoric acid buffer of 0.04-0.06mol/L pH6.8-7.2 in the triangular flask filling epoxidation composite nano fiber, under the temperature condition of 10-15 DEG C, with the velocity fluctuation 20-24h of 140-160r/min, prepared by immobilization TreP, clean up rear for subsequent use after taking out this immobilized enzyme by purified water.
Preferred according to the present invention, it is characterized in that, in described step (2), take high maltose syrup as substrate, utilize immobilization TreP to produce the step of trehalose as follows: with solid content 40-60%(w/w) high maltose syrup be substrate, get immobilization TreP with high maltose syrup with 1:(18-22) ratio of (w/w) mixes, immobilization TreP transforms the temperature of reaction of producing trehalose and controls as 30-40 DEG C, pH value in reaction is 4.0-6.5, reaction times is 18-60h, during content of trehalose >=55%, reaction terminates.
Preferred according to the present invention, in described step (2), the maltose content of high maltose syrup substrate used accounts for the 85-90%(w/w of solid content).
Preferred according to the present invention, in described step (3), double-enzyme composite hydrolysis method of purification adopts compounded saccharifying enzyme and the series connection of glucose transglucosidase to use.
Preferred according to the present invention, in described step (3), double-enzyme composite hydrolysis method of purification is adopted to improve the step of trehalose purity as follows: the trehalose saccharified liquid prepared according to step (2) to be carried out high temperature and go out after ferment treatment, feed liquid is diluted to concentration 35%-40% and is cooled to 58-60 DEG C, adjusting pH value of saccharified liquid is after 4.0-4.5, compounded saccharifying enzyme is added by the addition of 0.5-0.6L/t, after insulation >=12h, pH value is adjusted to 5.0-5.5, glucose transglucosidase is added again, insulation >=15h by the addition of 0.5-0.6L/t.When maltose after reaction to be detected in liquid glucose and isomaltose content are 0, carry out feed liquid high temperature and to go out ferment treatment, after process, the main component of feed liquid is glucose and trehalose.
Preferred according to the present invention, in described step (3), the high temperature ferment treatment that goes out is that feed liquid is warming up to 80-85 DEG C, insulation 20-25min.
Preferred according to the present invention, in described step (4), sugar component step more than trehalose, glucose and trisaccharide in the mixing solutions adopting chromatogram ternary component isolation technique division step (3) to obtain is as follows:
To according to step (3) process after feed liquid in add 0.1%(w/w) activated carbon carry out desolventing technology, decolouring 20-30min after, filter be purified rear feed liquid.This feed liquid processed through ion exchange resin, the metal ion in removing feed liquid and negatively charged ion, obtain the saccharified liquid that specific conductivity is less than 50 μ S/cm, then be the liquid glucose of 60-65% through concentrated acquisition concentration.This liquid glucose is separated through ternary separation simulation mobile bed chromatic, by sugar component more than trehalose, glucose and trisaccharide separately, thus improve the purity of trehalose and sugar component more than glucose and trisaccharide is reclaimed, a kind of moisture-keeping syrup of coproduction and glucose while production trehalose product.
Preferred according to the present invention, in described step (4), the controling parameters of chromatogram ternary component isolation technique comprises: ternary component separation simulation mobile bed chromatic separation temperature is 60-65 DEG C, the weight ratio of purified water and feed liquid is (2.0-2.2): 1, and treatment capacity is 0.02-0.022L/min.
Preferred according to the present invention, the component of the moisture-keeping syrup finally obtained is trehalose 75-78%, and the above component of glucose, trisaccharide and trisaccharide amounts to 22-25%.
the beneficial features of this technology trehalose project is:
(1) immobilized trehalose synthase can repeatedly use, and access times, more than 100 times, substantially increase the service efficiency of enzyme, makes Products production cost reduce 300 yuan/ton, for enterprise creates fairly high profit;
(2) eliminate the maltose affecting trehalose crystallization and isomaltose, primary crystallization can obtain the product that purity is greater than 99%, drastically increases the product quality of trehalose;
(3) a kind of moisture-keeping syrup is produced while preparing trehalose, without the generation of other byproducts in production process;
(4) three chromatographic separation technologies once realize the separation of three kinds of components, substitute the mode of 2 times original two kinds of Component seperation, easy to operate, while preparation high purity trehalose, obtain a kind of moisture-keeping syrup, the efficient preparation of whole process implementation trehalose and the comprehensive utilization of parting liquid.
Embodiment
Below in conjunction with embodiment, technical solution of the present invention is further elaborated.
Embodiment 1
A production method for crystalline trehalose coproduction moisture-keeping syrup, comprises the steps:
(1) adopt epoxidation composite nano fiber to the process of being fixed of TreP, concrete steps comprise:
Getting enzyme lives as the TreP enzyme liquid 90ml of 800U/ml, add after being diluted to 300ml with the phosphoric acid buffer of 0.04mol/L pH6.8 in the triangular flask filling epoxidation composite nano fiber, under the temperature condition of 15 DEG C, to shake 24h with the speed of 140r/min, prepared by immobilization TreP, clean up rear for subsequent use after taking out this immobilized enzyme by purified water.
(2) be substrate with high maltose syrup, the concrete steps utilizing immobilization TreP to produce trehalose comprise:
With solid content 60%(w/w), maltose content accounts for the 90%(w/w of solid content) high maltose syrup be substrate, get immobilization TreP with high maltose syrup with 1:18(w/w) ratio mix, immobilization TreP transforms the temperature of reaction of producing trehalose and controls to be 40 DEG C, pH value in reaction is 4.5, reaction times is 18h, during content of trehalose 55.6%, reaction terminates.
(3) double-enzyme composite hydrolysis method of purification is adopted, the trehalose obtain step (2) and the mixing solutions of maltose carry out purification process, remove the maltose and the isomaltose that affect trehalose crystallization purity, improve the purity of trehalose crystalline product, concrete steps are as follows:
The trehalose saccharified liquid prepared according to step (2) is warming up to 80 DEG C, and insulation 20min, carries out high temperature and to go out ferment treatment.Feed liquid after enzyme of going out is diluted to concentration 40% and is cooled to 60 DEG C, and adjusting pH value of saccharified liquid is after 4.5, adds compounded saccharifying enzyme by the addition of 0.5L/t, after insulation 12h, pH value is adjusted to 5.5, then adds glucose transglucosidase by the addition of 0.5L/t, insulation 15h.When maltose after reaction to be detected in liquid glucose and isomaltose content are 0, feed liquid is warming up to 85 DEG C, insulation 25min, then carries out feed liquid high temperature and to go out ferment treatment, after process, the main component of feed liquid is glucose and trehalose.
(4) sugar component more than trehalose, glucose and trisaccharide in the mixing solutions adopting chromatogram ternary component isolation technique division step (3) to obtain, object improves the purity of trehalose, reclaim the sugar component of more than glucose and trisaccharide, a kind of moisture-keeping syrup of coproduction while production trehalose product, concrete steps are as follows: to according to step (3) process after feed liquid in add 0.1%(w/w) activated carbon carry out desolventing technology, after decolouring 20min, filter and be purified rear feed liquid.This feed liquid processed through ion exchange resin, the metal ion in removing feed liquid and negatively charged ion, obtain the saccharified liquid of specific conductivity 50 μ S/cm, then be the liquid glucose of 60% through concentrated acquisition concentration.This liquid glucose is separated through ternary separation simulation mobile bed chromatic, ternary component separation simulation mobile bed chromatic separation temperature controls at 60 DEG C, the weight ratio of purified water and feed liquid controls at 2.0:1, treatment capacity controls at 0.02L/min, by sugar component more than trehalose, glucose and trisaccharide separately.The component of the moisture-keeping syrup finally obtained is trehalose 78%, and the above component of glucose, trisaccharide and trisaccharide amounts to 22%.
| Feed liquid situation after being separated | Glucose | Trehalose | More than trisaccharide and trisaccharide |
| Marine alga liquid glucose | 1.4% | 93.5% | 5.1% |
| Moisture-keeping syrup | 0 | 78.0% | 22.0% |
| Glucose Liquid | 98.6% | 0.3% | 1.1% |
Embodiment 2
A production method for crystalline trehalose coproduction moisture-keeping syrup, comprises the steps:
(1) adopt epoxidation composite nano fiber to the process of being fixed of TreP, concrete steps comprise:
Getting enzyme lives as the TreP enzyme liquid 100ml of 1000U/ml, add after being diluted to 290ml with the phosphoric acid buffer of 0.04mol/L pH6.8 in the triangular flask filling epoxidation composite nano fiber, under the temperature condition of 12 DEG C, with the velocity fluctuation 20h of 140r/min, prepared by immobilization TreP, clean up rear for subsequent use after taking out this immobilized enzyme by purified water.
(2) be substrate with high maltose syrup, the concrete steps utilizing immobilization TreP to produce trehalose comprise:
With solid content 45%(w/w), maltose content accounts for the 90%(w/w of solid content) high maltose syrup be substrate, get immobilization TreP with high maltose syrup with 1:18(w/w) ratio mix, immobilization TreP transforms the temperature of reaction of producing trehalose and controls to be 38 DEG C, pH value in reaction is 4.8, reaction times is 24h, during content of trehalose 56%, reaction terminates.
(3) double-enzyme composite hydrolysis method of purification is adopted, the trehalose obtain step (2) and the mixing solutions of maltose carry out purification process, remove the maltose and the isomaltose that affect trehalose crystallization purity, improve the purity of trehalose crystalline product, concrete steps are as follows:
The trehalose saccharified liquid prepared according to step (2) is warming up to 85 DEG C, and insulation 20min, carries out high temperature and to go out ferment treatment.Feed liquid after enzyme of going out is diluted to concentration 40% and is cooled to 58 DEG C, and adjusting pH value of saccharified liquid is after 4.5, adds compounded saccharifying enzyme by the addition of 0.55L/t, after insulation 13h, pH value is adjusted to 5.0, then adds glucose transglucosidase by the addition of 0.55L/t, insulation 16h.When maltose after reaction to be detected in liquid glucose and isomaltose content are 0, feed liquid is warming up to 85 DEG C, insulation 20min, then carries out feed liquid high temperature and to go out ferment treatment, after process, the main component of feed liquid is glucose and trehalose.
(4) sugar component more than trehalose, glucose and trisaccharide in the mixing solutions adopting chromatogram ternary component isolation technique division step (3) to obtain, object improves the purity of trehalose, reclaim the sugar component of more than glucose and trisaccharide, a kind of moisture-keeping syrup of coproduction while production trehalose product, concrete steps are as follows: to according to step (3) process after feed liquid in add 0.1%(w/w) activated carbon carry out desolventing technology, after decolouring 20min, filter and be purified rear feed liquid.This feed liquid processed through ion exchange resin, the metal ion in removing feed liquid and negatively charged ion, obtain the saccharified liquid of specific conductivity 45 μ S/cm, then be the liquid glucose of 62% through concentrated acquisition concentration.This liquid glucose is separated through ternary separation simulation mobile bed chromatic, ternary component separation simulation mobile bed chromatic separation temperature controls at 62 DEG C, the weight ratio of purified water and feed liquid controls at 2.1:1, treatment capacity controls at 0.021L/min, by sugar component more than trehalose, glucose and trisaccharide separately.The component of the moisture-keeping syrup finally obtained is trehalose 76.7%, and the above component of glucose, trisaccharide and trisaccharide amounts to 23.3%.
| Feed liquid situation after being separated | Glucose | Trehalose | More than trisaccharide and trisaccharide |
| Marine alga liquid glucose | 0.7% | 95.2% | 4.1% |
| Moisture-keeping syrup | 0 | 76.7% | 23.3% |
| Glucose Liquid | 98.9% | 0.3% | 0.8% |
Embodiment 3
A production method for crystalline trehalose coproduction moisture-keeping syrup, comprises the steps:
(1) adopt epoxidation composite nano fiber to the process of being fixed of TreP, concrete steps comprise:
Getting enzyme lives as the TreP enzyme liquid 100ml of 1200U/ml, add after being diluted to 300ml with the phosphoric acid buffer of 0.05mol/L pH7.0 in the triangular flask filling epoxidation composite nano fiber, under the temperature condition of 14 DEG C, with the velocity fluctuation 24h of 150r/min, prepared by immobilization TreP, clean up rear for subsequent use after taking out this immobilized enzyme by purified water.
(2) be substrate with high maltose syrup, the concrete steps utilizing immobilization TreP to produce trehalose comprise:
With solid content 50%(w/w), maltose content accounts for the 85%(w/w of solid content) high maltose syrup be substrate, get immobilization TreP with high maltose syrup with 1:20(w/w) ratio mix, immobilization TreP transforms the temperature of reaction of producing trehalose and controls to be 35 DEG C, pH value in reaction is 5.0, reaction times is 36h, during content of trehalose 56.5%, reaction terminates.
(3) double-enzyme composite hydrolysis method of purification is adopted, the trehalose obtain step (2) and the mixing solutions of maltose carry out purification process, remove the maltose and the isomaltose that affect trehalose crystallization purity, improve the purity of trehalose crystalline product, concrete steps are as follows:
The trehalose saccharified liquid prepared according to step (2) is warming up to 82 DEG C, and insulation 22min, carries out high temperature and to go out ferment treatment.Feed liquid after enzyme of going out is diluted to concentration 38% and is cooled to 58 DEG C, and adjusting pH value of saccharified liquid is after 4.3, adds compounded saccharifying enzyme by the addition of 0.58L/t, after insulation 16h, pH value is adjusted to 5.3, then adds glucose transglucosidase by the addition of 0.58L/t, insulation 18h.When maltose after reaction to be detected in liquid glucose and isomaltose content are 0, feed liquid is warming up to 82 DEG C, insulation 22min, then carries out feed liquid high temperature and to go out ferment treatment, after process, the main component of feed liquid is glucose and trehalose.
(4) sugar component more than trehalose, glucose and trisaccharide in the mixing solutions adopting chromatogram ternary component isolation technique division step (3) to obtain, object improves the purity of trehalose, reclaim the sugar component of more than glucose and trisaccharide, a kind of moisture-keeping syrup of coproduction while production trehalose product, concrete steps are as follows: to according to step (3) process after feed liquid in add 0.1%(w/w) activated carbon carry out desolventing technology, after decolouring 25min, filter and be purified rear feed liquid.This feed liquid processed through ion exchange resin, the metal ion in removing feed liquid and negatively charged ion, obtain the saccharified liquid of specific conductivity 30 μ S/cm, then be the liquid glucose of 63% through concentrated acquisition concentration.This liquid glucose is separated through ternary separation simulation mobile bed chromatic, ternary component separation simulation mobile bed chromatic separation temperature controls at 63 DEG C, the weight ratio of purified water and feed liquid controls at 2.0:1, treatment capacity controls at 0.02L/min, by sugar component more than trehalose, glucose and trisaccharide separately.The component of the moisture-keeping syrup finally obtained is trehalose 77.8%, and the above component of glucose, trisaccharide and trisaccharide amounts to 22.2%.
| Feed liquid situation after being separated | Glucose | Trehalose | More than trisaccharide and trisaccharide |
| Marine alga liquid glucose | 0.8% | 94.3% | 4.9% |
| Moisture-keeping syrup | 0 | 77.8% | 22.2% |
| Glucose Liquid | 99.1% | 0.2% | 0.7% |
Embodiment 4
A production method for crystalline trehalose coproduction moisture-keeping syrup, comprises the steps:
(1) adopt epoxidation composite nano fiber to the process of being fixed of TreP, concrete steps comprise:
Getting enzyme lives as the TreP enzyme liquid 110ml of 1500U/ml, add after being diluted to 310ml with the phosphoric acid buffer of 0.06mol/L pH7.2 in the triangular flask filling epoxidation composite nano fiber, under the temperature condition of 15 DEG C, with the velocity fluctuation 24h of 160r/min, prepared by immobilization TreP, clean up rear for subsequent use after taking out this immobilized enzyme by purified water.
(2) be substrate with high maltose syrup, the concrete steps utilizing immobilization TreP to produce trehalose comprise:
With solid content 60%(w/w), maltose content accounts for the 90%(w/w of solid content) high maltose syrup be substrate, get immobilization TreP with high maltose syrup with 1:22(w/w) ratio mix, immobilization TreP transforms the temperature of reaction of producing trehalose and controls to be 40 DEG C, pH value in reaction is 6.5, reaction times is 60h, during content of trehalose 58%, reaction terminates.
(3) double-enzyme composite hydrolysis method of purification is adopted, the trehalose obtain step (2) and the mixing solutions of maltose carry out purification process, remove the maltose and the isomaltose that affect trehalose crystallization purity, improve the purity of trehalose crystalline product, concrete steps are as follows:
The trehalose saccharified liquid prepared according to step (2) is warming up to 85 DEG C, and insulation 25min, carries out high temperature and to go out ferment treatment.Feed liquid after enzyme of going out is diluted to concentration 40% and is cooled to 60 DEG C, and adjusting pH value of saccharified liquid is after 4.5, adds compounded saccharifying enzyme by the addition of 0.6L/t, after insulation 18h, pH value is adjusted to 5.5, then adds glucose transglucosidase by the addition of 0.6L/t, insulation 20h.When maltose after reaction to be detected in liquid glucose and isomaltose content are 0, feed liquid is warming up to 85 DEG C, insulation 25min, then carries out feed liquid high temperature and to go out ferment treatment, after process, the main component of feed liquid is glucose and trehalose.
(4) sugar component more than trehalose, glucose and trisaccharide in the mixing solutions adopting chromatogram ternary component isolation technique division step (3) to obtain, object improves the purity of trehalose, reclaim the sugar component of more than glucose and trisaccharide, a kind of moisture-keeping syrup of coproduction while production trehalose product, concrete steps are as follows: to according to step (3) process after feed liquid in add 0.1%(w/w) activated carbon carry out desolventing technology, after decolouring 30min, filter and be purified rear feed liquid.This feed liquid processed through ion exchange resin, the metal ion in removing feed liquid and negatively charged ion, obtain the saccharified liquid of specific conductivity 25 μ S/cm, then be the liquid glucose of 65% through concentrated acquisition concentration.This liquid glucose is separated through ternary separation simulation mobile bed chromatic, ternary component separation simulation mobile bed chromatic separation temperature controls at 65 DEG C, the weight ratio of purified water and feed liquid controls at 2.2:1, treatment capacity controls at 0.022L/min, by sugar component more than trehalose, glucose and trisaccharide separately.The component of the moisture-keeping syrup finally obtained is trehalose 77.5%, and the above component of glucose, trisaccharide and trisaccharide amounts to 22.5%.
| Feed liquid situation after being separated | Glucose | Trehalose | More than trisaccharide and trisaccharide |
| Marine alga liquid glucose | 0.9% | 95.5% | 3.6% |
| Moisture-keeping syrup | 0 | 77.5% | 22.5% |
| Glucose Liquid | 99.5% | 0 | 0.5% |
Should be appreciated that shown in the invention is not restricted to here and the specific embodiment described.Those of ordinary skill in the art's all apparent change that can accomplish and changing on the basis of reading specification sheets of the present invention is all included in the scope of the present invention that appended claims limits.
Claims (10)
1. a production method for crystalline trehalose coproduction moisture-keeping syrup, is characterized in that, comprises the steps:
(1) epoxidation composite nano fiber is adopted to process being fixed of TreP;
(2) be substrate with high maltose syrup, utilize immobilization TreP to produce trehalose, when content of trehalose reaches 55%, terminate this reaction;
(3) adopt double-enzyme composite hydrolysis method of purification, the trehalose obtain step (2) and the mixing solutions of maltose carry out purification process, remove the maltose and the isomaltose that affect trehalose crystallization purity, improve the purity of trehalose crystalline product;
(4) sugar component more than trehalose, glucose and trisaccharide in the mixing solutions adopting chromatogram ternary component isolation technique division step (3) to obtain, object improves the purity of trehalose, reclaim the sugar component of more than glucose and trisaccharide, a kind of moisture-keeping syrup of coproduction while production trehalose product.
2. the method for claim 1, is characterized in that, in described step (1), adopts the step of epoxidation composite nano fiber to being fixed of TreP as follows:
Getting enzyme lives as the TreP enzyme liquid 90-110ml of 800-1500U/ml, add after being diluted to 290-310ml with the phosphoric acid buffer of 0.04-0.06mol/L pH6.8-7.2 in the triangular flask filling epoxidation composite nano fiber, under the temperature condition of 10-15 DEG C, with the velocity fluctuation 20-24h of 140-160r/min, prepared by immobilization TreP, clean up rear for subsequent use after taking out this immobilized enzyme by purified water.
3. the method for claim 1, is characterized in that, in described step (2), is substrate with high maltose syrup, utilizes the step of immobilization TreP production trehalose as follows:
With solid content 40-60%(w/w) high maltose syrup be substrate, get immobilization TreP with high maltose syrup with 1:(18-22) ratio of (w/w) mixes, immobilization TreP transforms the temperature of reaction of producing trehalose and controls as 30-40 DEG C, pH value in reaction is 4.0-6.5, reaction times is 18-60h, during content of trehalose >=55%, reaction terminates.
4. method as claimed in claim 3, it is characterized in that, the maltose content of high maltose syrup substrate used accounts for the 85-90%(w/w of solid content).
5. the method for claim 1, is characterized in that, in described step (3), double-enzyme composite hydrolysis method of purification adopts compounded saccharifying enzyme and the series connection of glucose transglucosidase to use.
6. the method for claim 1, is characterized in that, in described step (3), adopts the step of double-enzyme composite hydrolysis method of purification raising trehalose purity as follows:
The trehalose saccharified liquid prepared according to step (2) being carried out high temperature goes out after ferment treatment, feed liquid is diluted to concentration 35%-40% and is cooled to 58-60 DEG C, adjusting pH value of saccharified liquid is after 4.0-4.5, compounded saccharifying enzyme is added by the addition of 0.5-0.6L/t, after insulation >=12h, pH value is adjusted to 5.0-5.5, then adds glucose transglucosidase by the addition of 0.5-0.6L/t, insulation >=15h; When maltose after detection reaction in liquid glucose and isomaltose content are 0, carry out feed liquid high temperature and to go out ferment treatment, after process, the main component of feed liquid is glucose and trehalose.
7. method as claimed in claim 6, is characterized in that, the described high temperature ferment treatment that goes out is that feed liquid is warming up to 80-85 DEG C, insulation 20-25min.
8. the method for claim 1, is characterized in that, in described step (4), sugar component step more than trehalose, glucose and trisaccharide in the mixing solutions adopting chromatogram ternary component isolation technique division step (3) to obtain is as follows:
To according to step (3) process after feed liquid in add 0.1%(w/w) activated carbon carry out desolventing technology, decolouring 20-30min after, filter be purified rear feed liquid; This feed liquid processed through ion exchange resin, the metal ion in removing feed liquid and negatively charged ion, obtain the saccharified liquid that specific conductivity is less than 50 μ S/cm, then be the liquid glucose of 60-65% through concentrated acquisition concentration; This liquid glucose is separated through ternary separation simulation mobile bed chromatic, by sugar component more than trehalose, glucose and trisaccharide separately, thus improve the purity of trehalose and sugar component more than glucose and trisaccharide is reclaimed, a kind of moisture-keeping syrup of coproduction and glucose while production trehalose product.
9. method according to claim 8, is characterized in that, the controling parameters of chromatogram ternary component isolation technique comprises:
Ternary component separation simulation mobile bed chromatic separation temperature is 60-65 DEG C, and the weight ratio of purified water and feed liquid is (2.0-2.2): 1, and treatment capacity is 0.02-0.022L/min.
10. method according to claim 8, is characterized in that, the component of the moisture-keeping syrup finally obtained is trehalose 75-78%, and the above component of glucose, trisaccharide and trisaccharide amounts to 22-25%.
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| CN106399426A (en) * | 2016-10-10 | 2017-02-15 | 长沙理工大学 | Method for producing trehalose by cadmium-polluted rice |
| CN115838390A (en) * | 2022-12-30 | 2023-03-24 | 保龄宝生物股份有限公司 | Method for preparing high-content 2' -fucosyllactose by utilizing three-component chromatographic separation technology |
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