CN101200750A - Erwinia rhapontici and application thereof in preparation of isomaltulose - Google Patents
Erwinia rhapontici and application thereof in preparation of isomaltulose Download PDFInfo
- Publication number
- CN101200750A CN101200750A CNA2007101907552A CN200710190755A CN101200750A CN 101200750 A CN101200750 A CN 101200750A CN A2007101907552 A CNA2007101907552 A CN A2007101907552A CN 200710190755 A CN200710190755 A CN 200710190755A CN 101200750 A CN101200750 A CN 101200750A
- Authority
- CN
- China
- Prior art keywords
- sucrose
- cell
- palatinose
- isomaltulose
- erwinia
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 241000556426 Erwinia rhapontici Species 0.000 title claims abstract description 9
- PVXPPJIGRGXGCY-TZLCEDOOSA-N 6-O-alpha-D-glucopyranosyl-D-fructofuranose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)C(O)(CO)O1 PVXPPJIGRGXGCY-TZLCEDOOSA-N 0.000 title abstract description 11
- 238000002360 preparation method Methods 0.000 title description 6
- 229930006000 Sucrose Natural products 0.000 claims abstract description 46
- 239000005720 sucrose Substances 0.000 claims abstract description 46
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims abstract description 45
- 108010047540 sucrose isomerase Proteins 0.000 claims abstract description 24
- 238000006243 chemical reaction Methods 0.000 claims abstract description 23
- 241000588698 Erwinia Species 0.000 claims abstract description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 14
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 11
- 229910017053 inorganic salt Inorganic materials 0.000 claims abstract description 9
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 8
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 6
- 239000008103 glucose Substances 0.000 claims abstract description 6
- 239000001963 growth medium Substances 0.000 claims abstract description 6
- 229930091371 Fructose Natural products 0.000 claims abstract description 5
- 239000005715 Fructose Substances 0.000 claims abstract description 5
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 claims abstract description 5
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims abstract description 4
- PVXPPJIGRGXGCY-DJHAAKORSA-N 6-O-alpha-D-glucopyranosyl-alpha-D-fructofuranose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@](O)(CO)O1 PVXPPJIGRGXGCY-DJHAAKORSA-N 0.000 claims description 42
- 210000004027 cell Anatomy 0.000 claims description 21
- 240000004980 Rheum officinale Species 0.000 claims description 16
- 235000008081 Rheum officinale Nutrition 0.000 claims description 16
- 238000000034 method Methods 0.000 claims description 14
- 230000001580 bacterial effect Effects 0.000 claims description 12
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 claims description 11
- 210000001822 immobilized cell Anatomy 0.000 claims description 7
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 6
- 230000002255 enzymatic effect Effects 0.000 claims description 6
- 239000012528 membrane Substances 0.000 claims description 5
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 claims description 4
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 claims description 4
- 240000008042 Zea mays Species 0.000 claims description 4
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 4
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 4
- 229940041514 candida albicans extract Drugs 0.000 claims description 4
- 230000010307 cell transformation Effects 0.000 claims description 4
- 235000005822 corn Nutrition 0.000 claims description 4
- 244000005700 microbiome Species 0.000 claims description 4
- 239000012138 yeast extract Substances 0.000 claims description 4
- 229920002101 Chitin Polymers 0.000 claims description 3
- 244000068988 Glycine max Species 0.000 claims description 3
- 235000010469 Glycine max Nutrition 0.000 claims description 3
- 239000001888 Peptone Substances 0.000 claims description 3
- 108010080698 Peptones Proteins 0.000 claims description 3
- 229920002472 Starch Polymers 0.000 claims description 3
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 claims description 3
- 239000004202 carbamide Substances 0.000 claims description 3
- 235000010418 carrageenan Nutrition 0.000 claims description 3
- 229920001525 carrageenan Polymers 0.000 claims description 3
- 238000011049 filling Methods 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 239000008107 starch Substances 0.000 claims description 3
- 235000019698 starch Nutrition 0.000 claims description 3
- 235000012343 cottonseed oil Nutrition 0.000 claims description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-M dihydrogenphosphate Chemical compound OP(O)([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-M 0.000 claims description 2
- 235000012054 meals Nutrition 0.000 claims description 2
- 150000003016 phosphoric acids Chemical class 0.000 claims description 2
- 235000015598 salt intake Nutrition 0.000 claims description 2
- 159000000000 sodium salts Chemical class 0.000 claims description 2
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 claims 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims 1
- 229940072056 alginate Drugs 0.000 claims 1
- 235000010443 alginic acid Nutrition 0.000 claims 1
- 229920000615 alginic acid Polymers 0.000 claims 1
- 239000011575 calcium Substances 0.000 claims 1
- 229910052791 calcium Inorganic materials 0.000 claims 1
- 238000011534 incubation Methods 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 abstract description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract description 4
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- 230000007062 hydrolysis Effects 0.000 abstract description 2
- 238000006460 hydrolysis reaction Methods 0.000 abstract description 2
- 229910052757 nitrogen Inorganic materials 0.000 abstract description 2
- 238000007086 side reaction Methods 0.000 abstract description 2
- 238000005119 centrifugation Methods 0.000 abstract 1
- 238000009776 industrial production Methods 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 14
- 102000004190 Enzymes Human genes 0.000 description 12
- 108090000790 Enzymes Proteins 0.000 description 12
- 241000894006 Bacteria Species 0.000 description 9
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 description 7
- 239000000661 sodium alginate Substances 0.000 description 7
- 235000010413 sodium alginate Nutrition 0.000 description 7
- 229940005550 sodium alginate Drugs 0.000 description 7
- 241001622809 Serratia plymuthica Species 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 230000009466 transformation Effects 0.000 description 6
- 230000008569 process Effects 0.000 description 5
- 208000007976 Ketosis Diseases 0.000 description 4
- 239000006227 byproduct Substances 0.000 description 4
- 150000002584 ketoses Chemical class 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 238000002156 mixing Methods 0.000 description 4
- 238000004321 preservation Methods 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 238000005303 weighing Methods 0.000 description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 239000002504 physiological saline solution Substances 0.000 description 3
- 230000035484 reaction time Effects 0.000 description 3
- 239000011734 sodium Substances 0.000 description 3
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000006555 catalytic reaction Methods 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 235000021472 generally recognized as safe Nutrition 0.000 description 2
- -1 hydroxyl isomaltulose Chemical compound 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000012856 packing Methods 0.000 description 2
- 150000002939 palatinoses Chemical class 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 238000011218 seed culture Methods 0.000 description 2
- 238000012807 shake-flask culturing Methods 0.000 description 2
- 206010000060 Abdominal distension Diseases 0.000 description 1
- 241000589155 Agrobacterium tumefaciens Species 0.000 description 1
- 235000016068 Berberis vulgaris Nutrition 0.000 description 1
- 241000335053 Beta vulgaris Species 0.000 description 1
- 241000219310 Beta vulgaris subsp. vulgaris Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 206010067715 Gastrointestinal sounds abnormal Diseases 0.000 description 1
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 description 1
- AYRXSINWFIIFAE-SCLMCMATSA-N Isomaltose Natural products OC[C@H]1O[C@H](OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O)[C@@H](O)[C@@H](O)[C@@H]1O AYRXSINWFIIFAE-SCLMCMATSA-N 0.000 description 1
- 241000588748 Klebsiella Species 0.000 description 1
- 241000410414 Klebsiella sp. LX3 Species 0.000 description 1
- 241001014264 Klebsiella variicola Species 0.000 description 1
- 241000588701 Pectobacterium carotovorum Species 0.000 description 1
- 241000586779 Protaminobacter Species 0.000 description 1
- 240000000111 Saccharum officinarum Species 0.000 description 1
- 235000007201 Saccharum officinarum Nutrition 0.000 description 1
- 241000607715 Serratia marcescens Species 0.000 description 1
- 102000013275 Somatomedins Human genes 0.000 description 1
- 241000194025 Streptococcus oralis Species 0.000 description 1
- 235000021536 Sugar beet Nutrition 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- LPQOADBMXVRBNX-UHFFFAOYSA-N ac1ldcw0 Chemical compound Cl.C1CN(C)CCN1C1=C(F)C=C2C(=O)C(C(O)=O)=CN3CCSC1=C32 LPQOADBMXVRBNX-UHFFFAOYSA-N 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000000721 bacterilogical effect Effects 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 238000010924 continuous production Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000006196 deacetylation Effects 0.000 description 1
- 238000003381 deacetylation reaction Methods 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000011143 downstream manufacturing Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 238000012215 gene cloning Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 235000012907 honey Nutrition 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000003914 insulin secretion Effects 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- DLRVVLDZNNYCBX-RTPHMHGBSA-N isomaltose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)C(O)O1 DLRVVLDZNNYCBX-RTPHMHGBSA-N 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- QWIZNVHXZXRPDR-WSCXOGSTSA-N melezitose Chemical compound O([C@@]1(O[C@@H]([C@H]([C@@H]1O[C@@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)O)CO)CO)[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O QWIZNVHXZXRPDR-WSCXOGSTSA-N 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 239000006916 nutrient agar Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000008057 potassium phosphate buffer Substances 0.000 description 1
- 230000002285 radioactive effect Effects 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M sodium chloride Inorganic materials [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 210000001082 somatic cell Anatomy 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 125000000185 sucrose group Chemical group 0.000 description 1
- 150000005846 sugar alcohols Chemical class 0.000 description 1
- 235000021092 sugar substitutes Nutrition 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
- 239000001052 yellow pigment Substances 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses an Erwinia rhapontici (Erwinia haryphontici) NX-5 and a method for producing sucrose isomerase by using the Erwinia rhapontici to prepare isomaltulose. The strain CGMCC No.2222 is inoculated in a sterile culture medium containing a carbon source, a nitrogen source, inorganic salt and water for aerobic culture, and cells containing sucrose isomerase are obtained by centrifugation or ultrafiltration, and then free cells containing sucrose isomerase are directly adopted to convert sucrose to generate isomaltulose or the cells containing sucrose isomerase are immobilized and then converted into sucrose to generate isomaltulose. The strain is adopted to ensure that the conversion rate of sucrose reaches 99.5% (w/w), the conversion rate of isomaltulose reaches 90%, the concentration of isomaltulose in the conversion solution reaches 500g/L, hydrolysis side reaction does not exist, the conversion solution hardly contains glucose and fructose, the product isomaltulose can not be converted into other components along with the reaction, and the strain is very beneficial to the industrial production of isomaltulose.
Description
Technical field
The invention belongs to fermentation engineering and technical field of enzyme engineering, relate to a kind of microorganism strains rheum officinale Erwinia (Erwinia rhapontici) NX-5 that produces sucrose isomerase, and it is used for the production of Palatinose.
Background technology
(6-O-α-D-glucopyranosyl-D-fructose Isomaltulose) is a kind of reducing sugar to Palatinose, exists on a small quantity in natural honey and sugar beet juice, has and the most similar physical properties of sucrose and mouthfeel.It has following advantage as the substitute of sucrose: (1) since it stimulated insulin secretion slowly and not than sucrose by the edible back of human body discharges monose in blood speed, can be edible for diabetic population and fat-reducing personage; (2) do not caused the Streptococcus oralis metabolism of carious tooth, do not corroded tooth; (3) in the micropopulation district of people's intestines, the growth of selective stimulating bifidus bacillus; (4) performance edible safety height is given food safety highest ranking " GRAS (generally recognized as safe) " by U.S. FDA, to its every day intake do not limit, can not make us producing untoward reactions such as abdominal distension, borborygmus.(5) have extremely low water absorbability, the hard candy of making of it can be as not becoming sticky with common sugar or other sugar substitutes, even can be packed by loosely.Because water absorbability is little, so stability is strong, shelf-lives is also longer.This is the difficult realizations of functional sugar alcohol such as Xylitol and maltose alcohol.(6) another major advantage of being approved by market of hydroxyl isomaltulose is a pure taste, any purposes that it almost can place of sucrose, and the human consumer can not differentiate and the difference that confectionery is arranged, and makes sugarfree foods " delicious food does not give a discount ".Because these characteristics, Palatinose (alcohol) become most popular sucrose substitute at present.
Palatinose is by sucrose isomerase EC 5.4.99.11 (Sucrose isomerase), or be Palatinose synthetic enzyme (Isomaltulose syntheses), sucrose glucosyl group mutase (Sucrose glucosylmutase), (α-glucosyltransferase) transformation of catalysis sucrose forms the alpha-glucosyl transferring enzyme, and known have some bacterial classifications can produce this enzyme.United States Patent (USP) 4,359,531 adopt the rheum officinale Erwinia to transform sucrose, and approximately the sucrose of 70-95% is transformed into Palatinose.United States Patent (USP) 4,390,627 have described the fixing sucrose mutase method of producing Palatinose from Protaminobacter rubrum (protaminobacter ruber).United States Patent (USP) 4,670,387 have described use Erwinia rhapontici, Protaminobacter rubrum, the immobilized thallus of Serratia plymuthica (serratia marcescens) is produced the process of Palatinose, approximately can transform 70-95% sucrose.United States Patent (USP) 4,857,461 disclose from Protaminobacterrubrum, the process of the thick enzyme continuous production of the immobilization Palatinose that Serratia plymuthica and Erwinia carotovora bacterium obtain.U.S. Patent No. 5,229,276 and No.5,336,617 have described the process of producing marine alga ketose and Palatinose with the immobilized thallus of Agrobacteriumradiobacter (radioactive soil bacillus), and Palatinose accounts for below 10%.Though above-mentioned several bacterium can be transformed into Palatinose to sucrose, output is very unstable, and transformation efficiency is 8～86%.And, these bacterial strains of producing Palatinose are in catalysis sucrose inversion process, except principal product, also exist marine alga ketose, isomaltose, different melizitose to reach glucose, the fructose by product that generates owing to sucrose hydrolysis in the enzymatic conversion liquid, the product specificity is not high, reduced the yield of purpose product on the one hand, made downstream process become very difficult on the other hand, production cost increases greatly.We produce bacterium to the Palatinose of different sources and are analyzed.It is few to find that rheum officinale Erwinia NX-5 CGMCC No.2222 transforms the by product that is generated, and is more conducive to further suitability for industrialized production, as shown in table 1.
Recently patent CN1434861 relates to two new bacterial strains, Singapore's klebsiella (Klebsiellasingaporensis) LX3 and LX21, there is higher enzyme to live, the content of Palatinose surpasses 87%, and the nucleotide sequence and this gene cloning and expression that disclose the novel type of sucrose isomerase make it generate Palatinose in plants such as sugarcane, corn, beets.Screening can the High-efficient Production Palatinose bacterial strain, the katalysis mechanism of further understanding enzyme is still very significant.
Summary of the invention
The rheum officinale Erwinia (Erwinia rhapontici) that the purpose of this invention is to provide a kind of high yield sucrose isomerase.
Another object of the present invention provides utilizes above-mentioned rheum officinale Erwinia to prepare the method for Palatinose.
The object of the invention can reach by following measures:
Microorganism strains rheum officinale Erwinia (Erwinia rhapontici) NX-5 of seed selection of inventor laboratory and preservation, this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center (being called for short CGMCC) at present, the numbering of registering on the books is CGMCC No.2222, and preservation date is: on October 22nd, 2007.With this bacterium as producing bacterial strain.
CGMCC No.2222 bacterial strain has following character:
(1) colonial morphology feature:
On nutrient agar medium, to cultivate 24h for 30 ℃ and faint yellow small colonies occurs, bacterium colony is rounded, smooth surface, sticking shape, center projections is opaque.Proper extension is cultivated, and bacterium colony increases, and the size and dimension of cell changes very little.
(2) physiology and biochemical characteristic:
A. culture temperature: 25～35 ℃, optimum temperuture is 30 ℃;
B. in pH 5～8 scopes, grow;
C. pigment produces: produce yellow pigment;
D. anti-NaCl concentration: can grow in 5% concentration.
(3) nutritional character:
Do not need to add somatomedin in the substratum of rheum officinale Erwinia NX-5.Organonitrogen or inorganic nitrogen can use as nitrogenous source.
The method that the present invention utilizes rheum officinale Erwinia CGMCC No.2222 to produce sucrose isomerase and then generation Palatinose is this inoculation have been carried out air culture support in the aseptic culture medium of carbonaceous sources, nitrogenous source, inorganic salt and water, centrifugal or ultrafiltration obtains to contain the cell of sucrose isomerase, and then the cell transformation sucrose that directly adopts free to contain sucrose isomerase generates and transforms sucrose behind the cell that Palatinose or immobilization contain sucrose isomerase and generate Palatinose.
Condition of enzyme production:
Rheum officinale Erwinia NX-5 CGMCC No.2222 slant activation after one day (the slant culture based component also contains 2% agar except that containing carbon source, nitrogenous source, inorganic salt etc.), is inoculated in the aseptic culture medium of carbonaceous sources, nitrogenous source, inorganic salt and water.Each components contents is percent weight in volume in the substratum, and promptly the g/100ml substratum is as follows.Ratio 2～the 10g/100ml of carbon source consumption and substratum, the ratio 0.2～5g/100ml of nitrogenous source consumption and substratum, the ratio 0.01～1g/100ml of inorganic salt consumption and substratum, all the other are water.The carbon source of substratum is one or more in glucose, sucrose, fructose, maltose, the starch hydrolyzate; Nitrogenous source is extractum carnis, peptone, yeast extract paste, corn steep liquor, soybean cake powder, cottonseed meal, urea, (NH
4)
2SO
4, NH
4Among the Cl one or more; Inorganic salt are one or more in sodium salt, phosphoric acid salt, the dihydrogen phosphate.The initial pH scope of substratum is 6.0～8.0,25～35 ℃ of leavening temperatures, shaking speed 100～200r/min, cultivate 8～20h, wet cell weight reaches 20g/L, and fermentation broth enzyme work reaches 2.5～5.0U/ml, centrifugal then or ultrafiltration obtains to contain the cell of sucrose isomerase, centrifugal condition: under the room temperature condition, 5000～10000r/min, 10～30min.Adopt the used ultra-filtration membrane of ultrafiltration process aperture 0.01～0.1 μ m, working pressure 0.1～0.6Mpa, the ultra-filtration membrane molecular weight that dams is 10
5～10
6Dalton.
Sucrose isomerase enzyme activity determination method:
The mixture that contains 40% (w/v) sucrose and 75g/L cell suspending liquid with potassium phosphate buffer (pH7.0) preparation of 0.1M, get this mixture 1ml behind 30 ℃ water-bath internal reaction 60min, the boiling water 10min that places 100 ℃ is cooled to room temperature with tap water then to stop enzyme reaction.Reaction mixture is measured the Palatinose growing amount through centrifugal and filtration back with the HPLC method.The sucrose isomerase enzyme work of a unit is defined as under 30 ℃, and pH is the enzyme amount that per minute generates the Palatinose of 1 μ mol under 7.0 the condition.
Enzymatic conversion method:
The cell that will contain sucrose isomerase carries out immobilization, process for fixation is: preparation 1～3% (the i.e. sodium alginate of 1～3g/100ml) concentration, the cell of adding 10～25% (being the cells that add 10～25 grams in every 100ml sodium alginate soln), and add 5～10% diatomite (being the diatomite that adds 5～10 grams in every 100ml sodium alginate soln), be 1～4% CaCl again with concentration
2Solution (g/100ml) is fixed-type with the embedded material sodium alginate, reacts as the enzyme source with this.Except sodium alginate as the fixation support embedding, can also use carrier embedding cells such as carrageenin, chitin.
Specifically: the cell that contains sucrose isomerase after will the dissociate cell that contains sucrose isomerase or the immobilization is filled in the retort, the sucrose solution that adds 450～600g/L, perhaps will pack in the filling type column type reactor according to the immobilized cell that the said fixing method makes, the volume of post is not limit.This reactor can 0.5～5ml/min way flow adds or the circulation Continuous Flow adds the sucrose solution of 450～600g/L, under 25～35 ℃ condition, carry out enzymatic conversion reaction, reaction times 10～15h, by adjusting flow velocity, the transformation efficiency of control sucrose is between 90～100%, collect effluent liquid, condensing crystal obtains Palatinose.
Beneficial effect of the present invention:
The present invention screens a kind of bacterial strain that can efficiently transform sucrose generation Palatinose, the sucrose inversion rate reaches as high as 99.5% (w/w), the Palatinose transformation efficiency can reach 90%, Palatinose concentration can reach 500g/L in the conversion fluid, nearly 10: 1 of Palatinose/marine alga ketose ratio, topmost do not have a hydrolytic side reactions, contain glucose and fructose in the conversion fluid hardly, in addition along with reaction is carried out also can not being converted into other compositions from the product Palatinose, these character be the bacterial strain in the past reported do not have, this is very useful to the suitability for industrialized production Palatinose.
Reference background technical literature material is produced bacterium to different Palatinoses and is analyzed, and the result is as shown in table 1:
The different Palatinoses of table 1. are produced the comparison of bacterium
| Bacterial strain | Optimal pH | Optimum temperuture (℃) | Palatinose/marine alga ketose ratio | By product ratio (%) |
| E.rhapontici S.plymuthica Klebsiella sp.LX3 CGMCC No.2222 | 7.0 6.0 6.0 7.0 | 30 30 35 30 | 5∶1～8∶1 8∶1 8∶1 10∶1 | 10.5 11.7 9.4 3.6 |
It is few to found that rheum officinale Erwinia NX-5 CGMCC No.2222 transforms the by product that is generated, and is more conducive to further suitability for industrialized production.
Embodiment
The invention will be further described by the following examples, but to the present invention without limits.
Embodiment 1
Slant medium: peptone 10g/L, extractum carnis 3g/L, NaCl5g/L, agar 20g/L, pH7.0.
Shake-flask culture base: sucrose 50g/L, yeast extract paste 10g/L, Na
3PO
412H
2O 5g/L, pH7.0.
Rheum officinale Erwinia NX-5 CGMCC No.2222 30 ℃ of cultivation 24h on slant medium, connect this bacterium of ring then in the shake-flask culture base, cultivate 10h for 30 ℃, shake a bottle rotating speed 200r/min, content of thalli is 20g/L in the fermented liquid that obtains, produce enzyme and reach 2.5～5.0U/ml, transform with free cell, the cell that takes by weighing 10g adds in the sucrose solution of 100ml450g/L (the cell addition is by the sucrose solution of every 100g cell transformation 1L55%), under 30 ℃ condition, vibration transforms and produces Palatinose in shaking bottle, and reaction times 8h surveys substrate conversion efficiency and production concentration after the stopping of reaction.The sucrose inversion rate is 99.5%, and the Palatinose transformation efficiency reaches 90%.
Embodiment 2
With 50g/L maltose is carbon source, and the 5g/L soybean cake powder is a nitrogenous source, and other composition of substratum and yeast culture condition are with embodiment 1, the sodium alginate colloidal solution of preparation 1.5%, after mixing with the synthetic bacteria suspension of physiological saline, add 8% diatomite adsorption after, splash into 2% CaCl with syringe
2Solidify in the solution, make the spherical immobilized cell that diameter is about 3～4mm, leave standstill a few hours in 4 ℃ of refrigerators, supernatant liquor then inclines, through distilled water wash 2 times, the 10g wet cell is obtained the 20g immobilized cell with sodium alginate to embed, under 30 ℃ condition, in shaking bottle, transform the sucrose solution of 100ml550g/L with this, every approving and forwarding time 10h, transform 40 batches, the average conversion that obtains sucrose is 99.5%, and the average conversion of Palatinose is 88%.
Embodiment 3
With glucose 225g, corn steep liquor 45g, NaCl22.5g, pH6.0, the water constant volume is made into substratum to 4.5L, in the 7.5 liters of glass stirred fermentors of packing into, 121 ℃ of steam sterilizing 15min.Preparation seed culture medium: sucrose 50g/L, yeast extract paste 10g/L, Na
2HPO
412H
2O 5g/L, pH6.0.Rheum officinale Erwinia NX-5 CGMCC No.2222 is connect one to be encircled in seed culture medium, cultivate 8h for 30 ℃ and get seed liquor, the inoculum size of seed liquor by fermentating liquid volume 3% inserted in the cooled fermention medium 30 ℃ of cultivations (ventilation 1vvm, mixing speed are 600r/min) 12h.Fermented liquid is carried out bacteriological filtration handle in ultra-fine filter, the ultra-filtration membrane molecular weight that dams is 10
6Dalton, working pressure is 0.2MPa, 50 ℃ of controlled temperature, face speed 4m/s, after the method immobilization of 90g thalline that obtains of will damming by embodiment 2, in the retort of 2L of packing into, add the sucrose solution 900ml of 550g/L, 30 ℃ of temperature of reaction, mixing speed 50r/min, reaction times 13h, the sucrose inversion rate is 99%, the Palatinose transformation efficiency reaches 85%.
Embodiment 4
With the 50g/L starch hydrolyzate is carbon source, and 20g/L urea is nitrogenous source, and other composition of substratum and yeast culture condition take by weighing the 160g wet thallus with embodiment 1, add sterilized water 100mL, in 30 ℃ of water bath heat preservations; Take by weighing the 10g carrageenin, add sterilized water 200mL, fully soak heating for dissolving.Both are mixed rapidly, pave plate, place 30min, be cut into the particle of size about 2*2*2 after solidifying, in 2mol/LKC1 solution, soak 30min again, it is fully solidified in 4 ℃ of refrigerators.Leach particle and wash 2-3 time with physiological saline, 4 ℃ of preservations are standby.The gained thalline forms immobilized cell 350g, pack in the filling bed type column type reactor of φ 4 * 45cm, (two placed in-line jacketed type immobilization posts), the dress post amount of immobilized cell is 80% (v/v), the sucrose solution that adds 550g/L with the flow velocity way flow of 1ml/min, carry out enzymatic conversion under 30 ℃ the condition, 15h is one batch.This device moves 30 days continuously, transforms sucrose solution 55L, and the average conversion that obtains sucrose is 99.5%, and the average conversion of Palatinose is 88%.
Embodiment 5
Yeast culture took by weighing the chitin of 30 mesh sieves with embodiment 1, at 80 ℃ of following deacetylations, was made into the aqueous solution by wet thallus content 2% with 40%NaOH then, regulated pH to subacidity with NaOH; Somatic cells and equivalent physiological saline mixing; With injection needle tube said mixture is splashed into 3% Na
3PO
4In the solution, in phosphate buffer solution, soak to solidify 24h, diameter be the bead of 2mm.The 400g immobilized spherule is loaded in the retort, adds the sucrose solution of 2L600g/L, carry out enzymatic conversion reaction by the condition of embodiment 3, transform 42 batches, the average conversion that obtains sucrose is 99%, and the average conversion of Palatinose is 85%.
Claims (8)
1. rheum officinale Erwinia NX-5, its classification called after rheum officinale Erwinia (Erwinia rhapontici) NX-5 has been preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, and its deposit number is: CGMCC No.2222.
2. the application of the described rheum officinale Erwinia of claim 1 in producing Palatinose.3, application according to claim 2, the concrete grammar that it is characterized in that this application is for cultivating in the aseptic culture medium that this bacterial strain CGMCC No.2222 is inoculated in carbonaceous sources, nitrogenous source, inorganic salt and water, centrifugal or ultrafiltration obtains to contain the cell of sucrose isomerase, and then the cell transformation sucrose that directly adopts free to contain sucrose isomerase generates and transforms sucrose behind the cell that Palatinose or immobilization contain sucrose isomerase and generate Palatinose.
4. application according to claim 3, it is characterized in that in the described substratum, each amounts of components is: the carbon source consumption is 2～10g/100ml, and the nitrogenous source consumption is 0.2～5g/100ml, and the inorganic salt consumption is 0.01～1g/100ml, and all the other are water.
5. according to claim 3 or 4 described application, the carbon source that it is characterized in that described substratum is one or more in glucose, sucrose, fructose, maltose, the starch hydrolyzate; Nitrogenous source is extractum carnis, peptone, yeast extract paste, corn steep liquor, soybean cake powder, cottonseed meal, urea, (NH
4)
2SO
4, NH
4Among the Cl one or more; Inorganic salt are one or more in sodium salt, phosphoric acid salt, the dihydrogen phosphate.
6. application according to claim 3, it is characterized in that bacterial strain CGMCC No.2222 is carried out culture condition is: the initial pH of substratum is 6.0～8.0, and culture temperature is 25～35 ℃, and incubation time is 8～20h.
7. application according to claim 3 is characterized in that adopting the method for centrifugal or ultrafiltration to obtain to contain the sucrose isomerase cell, and used ultra-filtration membrane aperture 0.01-0.1 μ m, working pressure 0.1-0.6Mpa, the ultra-filtration membrane molecular weight that dams is 10
5～10
6Dalton.
8. application according to claim 3, it is that the cell after free cell or the immobilization is filled in the retort that the cell transformation sucrose that it is characterized in that containing sucrose isomerase is produced Palatinose, adds the sucrose solution of 450～600g/L; Perhaps immobilized cell is packed in the filling bed type column type reactor, this reactor adds with the flow velocity way flow of 0.5～5ml/min or the circulation Continuous Flow adds the sucrose solution of 450～600g/L, carry out enzymatic conversion reaction, 25～35 ℃ of temperature of reaction, every approving and forwarding time 8～15h.
9. application according to claim 8 is characterized in that immobilized cell is to adopt alginate calcium, carrageenin or chitin as fixation support.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007101907552A CN101200750B (en) | 2007-11-29 | 2007-11-29 | Erwinia rhapontici and application thereof in preparation of isomaltulose |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007101907552A CN101200750B (en) | 2007-11-29 | 2007-11-29 | Erwinia rhapontici and application thereof in preparation of isomaltulose |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101200750A true CN101200750A (en) | 2008-06-18 |
| CN101200750B CN101200750B (en) | 2010-09-29 |
Family
ID=39516100
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2007101907552A Expired - Fee Related CN101200750B (en) | 2007-11-29 | 2007-11-29 | Erwinia rhapontici and application thereof in preparation of isomaltulose |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101200750B (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102286454A (en) * | 2011-07-01 | 2011-12-21 | 广西科学院生物研究所 | Method for preparing isomaltulose by polyvinyl alcohol immobilized sucrose isomerase producing bacteria |
| CN104017028A (en) * | 2014-06-23 | 2014-09-03 | 齐鲁工业大学 | Method of separating isomaltulose and trehalulose from isomaltulose mother liquor |
| CN105378097A (en) * | 2013-07-18 | 2016-03-02 | 甜糖（曼海姆/奥克森富特）股份公司 | Optimised method for producing a composition containing isomaltulose |
| CN105363039A (en) * | 2014-09-01 | 2016-03-02 | 中国海洋大学 | Enteromorpha polysaccharide microcapsule and preparation method thereof |
| CN105907818A (en) * | 2016-06-17 | 2016-08-31 | 青岛哈尼康生物科技有限公司 | Isomaltulose production process |
| CN105969824A (en) * | 2016-07-25 | 2016-09-28 | 山东百龙创园生物科技有限公司 | Preparation method of palatinitol |
| CN110172488A (en) * | 2019-06-13 | 2019-08-27 | 赵春海 | A kind of preparation process producing isomaltoketose |
| CN112063666A (en) * | 2020-08-05 | 2020-12-11 | 浙江工业大学 | Application of recombinant sucrose isomerase in preparation of isomaltulose by converting sucrose |
| CN115029338A (en) * | 2022-05-31 | 2022-09-09 | 黄凯奇 | Method for producing isomaltulose by fermenting sucrose isomerase |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1330149A (en) * | 2000-06-20 | 2002-01-09 | 范志刚 | Process for preparing isomaltoketose |
| CN100396780C (en) * | 2005-06-22 | 2008-06-25 | 万代生物技术（深圳）有限公司 | Process for preparing isomaltitol |
-
2007
- 2007-11-29 CN CN2007101907552A patent/CN101200750B/en not_active Expired - Fee Related
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102286454A (en) * | 2011-07-01 | 2011-12-21 | 广西科学院生物研究所 | Method for preparing isomaltulose by polyvinyl alcohol immobilized sucrose isomerase producing bacteria |
| CN102286454B (en) * | 2011-07-01 | 2013-04-17 | 广西科学院生物研究所 | Method for preparing isomaltulose by polyvinyl alcohol immobilized sucrose isomerase producing bacteria |
| CN105378097B (en) * | 2013-07-18 | 2019-06-21 | 甜糖（曼海姆/奥克森富特）股份公司 | Prepare the optimization method of the composition containing isomaltoketose |
| CN105378097A (en) * | 2013-07-18 | 2016-03-02 | 甜糖（曼海姆/奥克森富特）股份公司 | Optimised method for producing a composition containing isomaltulose |
| CN104017028B (en) * | 2014-06-23 | 2016-06-29 | 齐鲁工业大学 | The method separating isomaltulose and trehalulose from isomaltulose mother solution |
| CN104017028A (en) * | 2014-06-23 | 2014-09-03 | 齐鲁工业大学 | Method of separating isomaltulose and trehalulose from isomaltulose mother liquor |
| CN105363039A (en) * | 2014-09-01 | 2016-03-02 | 中国海洋大学 | Enteromorpha polysaccharide microcapsule and preparation method thereof |
| CN105363039B (en) * | 2014-09-01 | 2018-09-28 | 中国海洋大学 | A kind of sea grass polysaccharide microcapsules and preparation method thereof |
| CN105907818A (en) * | 2016-06-17 | 2016-08-31 | 青岛哈尼康生物科技有限公司 | Isomaltulose production process |
| CN105969824A (en) * | 2016-07-25 | 2016-09-28 | 山东百龙创园生物科技有限公司 | Preparation method of palatinitol |
| CN110172488A (en) * | 2019-06-13 | 2019-08-27 | 赵春海 | A kind of preparation process producing isomaltoketose |
| CN112063666A (en) * | 2020-08-05 | 2020-12-11 | 浙江工业大学 | Application of recombinant sucrose isomerase in preparation of isomaltulose by converting sucrose |
| CN112063666B (en) * | 2020-08-05 | 2021-10-15 | 浙江工业大学 | Application of recombinant sucrose isomerase in preparation of isomaltulose by converting sucrose |
| CN115029338A (en) * | 2022-05-31 | 2022-09-09 | 黄凯奇 | Method for producing isomaltulose by fermenting sucrose isomerase |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101200750B (en) | 2010-09-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101200750B (en) | Erwinia rhapontici and application thereof in preparation of isomaltulose | |
| KR970009295B1 (en) | Process for preparing trehalulose and isomaltulose | |
| CN102492673A (en) | Method for producing alternan sucrase by fermenting Leuconostoccitreum and its application | |
| CN102296032B (en) | Transglucosidase, its preparation method and immobilization method | |
| CN101531975B (en) | Lactobacillus fermentum and method for preparing D-tagatose by using same | |
| CN101974455B (en) | Escherichia coli strain for high yield of Gamma-aminobutyric acid and method for producing Gamma-aminobutyric acid therefrom | |
| CN101857890A (en) | Method for biologically converting stevioside in stevia sugar into rebaudioside | |
| CN105219663A (en) | The special strain therefore of trehalose synthesis and the method for the synthesis of trehalose thereof | |
| CN101486984B (en) | Gluconobacter oxydans and method for preparing ketoxylose using the same | |
| CN102690762A (en) | Method for producing L-alanine by bioconverting L-aspartic acid by free cell process | |
| CN1231593C (en) | Technology for producing glucomannan using neutral beta-mannase to degradate fine konjaku flour | |
| CN102127515B (en) | Screening and application of L-proline high-producing Brevundimonas sp. (JNPP-1) | |
| CN102559538A (en) | Escherichia.coli with high L-aspartase yield and application thereof | |
| CN106987573A (en) | It is a kind of to produce malt oligosaccharide based mycose synthetase, the method for malt oligosaccharide based mycose hydrolase and the method for producing trehalose | |
| KR102013010B1 (en) | Strain producing turanose and uses thereof | |
| CN102533607B (en) | Strain capable of producing beta-galactosidase and method for producing galactooligosaccharides by using beta-galactosidase | |
| CN1151253C (en) | Mycose base hydrolase and its preparation and use | |
| CN100526470C (en) | Preparation method of functional sweetener D-tatai sugar | |
| CN103045561B (en) | Solid state fermentation production method for beta-D-fructofuranosidase | |
| JP4338356B2 (en) | Microorganism producing fructosyltransferase, and method for producing fructooligosaccharide and neoflactooligosaccharide using the same | |
| CN102286569B (en) | Curdlan and preparation method thereof | |
| CN102229970B (en) | Method for producing dextran and fructose by recombinant enzyme process | |
| CN105861344A (en) | Synchronous culture method for improving yeast biomass and intracellular trehalose content | |
| CN85104280A (en) | A kind of method of utilizing microorganism continuous brewing water fruit vinegar | |
| CN115058352B (en) | Bacillus subtilis and method for producing agricultural gamma-polyglutamic acid by using same |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20100929 Termination date: 20211129 |