CN105087716B - A kind of sweet potato waste rapid enzymolysis at monosaccharide method - Google Patents
A kind of sweet potato waste rapid enzymolysis at monosaccharide method Download PDFInfo
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- CN105087716B CN105087716B CN201410198435.1A CN201410198435A CN105087716B CN 105087716 B CN105087716 B CN 105087716B CN 201410198435 A CN201410198435 A CN 201410198435A CN 105087716 B CN105087716 B CN 105087716B
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- 244000017020 Ipomoea batatas Species 0.000 title claims abstract description 59
- 235000002678 Ipomoea batatas Nutrition 0.000 title claims abstract description 59
- 239000002699 waste material Substances 0.000 title claims abstract description 39
- 238000000034 method Methods 0.000 title claims abstract description 9
- 150000002772 monosaccharides Chemical class 0.000 title claims description 9
- 229920002472 Starch Polymers 0.000 claims abstract description 19
- 239000008107 starch Substances 0.000 claims abstract description 19
- 235000019698 starch Nutrition 0.000 claims abstract description 19
- 108090000790 Enzymes Proteins 0.000 claims abstract description 17
- 102000004190 Enzymes Human genes 0.000 claims abstract description 17
- 238000000855 fermentation Methods 0.000 claims abstract description 17
- 239000001913 cellulose Substances 0.000 claims abstract description 16
- 229920002678 cellulose Polymers 0.000 claims abstract description 16
- 239000007788 liquid Substances 0.000 claims abstract description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 241000894006 Bacteria Species 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 5
- 239000002893 slag Substances 0.000 claims description 5
- 241000987688 Caldicellulosiruptor kronotskyensis Species 0.000 claims description 4
- 241000511679 Caldicellulosiruptor lactoaceticus Species 0.000 claims description 4
- 239000013589 supplement Substances 0.000 claims description 4
- 241000178334 Caldicellulosiruptor Species 0.000 claims description 3
- 239000000411 inducer Substances 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 2
- 230000004151 fermentation Effects 0.000 abstract description 15
- 229940088598 enzyme Drugs 0.000 abstract description 14
- 239000000843 powder Substances 0.000 abstract description 10
- 230000007071 enzymatic hydrolysis Effects 0.000 abstract description 8
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 abstract description 8
- 108010059892 Cellulase Proteins 0.000 abstract description 6
- 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 abstract description 6
- 229940106157 cellulase Drugs 0.000 abstract description 6
- 239000008103 glucose Substances 0.000 abstract description 6
- 235000013325 dietary fiber Nutrition 0.000 abstract description 2
- 239000000284 extract Substances 0.000 abstract description 2
- 230000007062 hydrolysis Effects 0.000 abstract description 2
- 238000006460 hydrolysis reaction Methods 0.000 abstract description 2
- 108090000623 proteins and genes Proteins 0.000 abstract description 2
- 102000004169 proteins and genes Human genes 0.000 abstract description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract 2
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 abstract 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 abstract 1
- 229910052799 carbon Inorganic materials 0.000 abstract 1
- 235000019441 ethanol Nutrition 0.000 abstract 1
- 239000004449 solid propellant Substances 0.000 abstract 1
- 235000010980 cellulose Nutrition 0.000 description 13
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 10
- 239000004382 Amylase Substances 0.000 description 6
- 108010065511 Amylases Proteins 0.000 description 6
- 102000013142 Amylases Human genes 0.000 description 6
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 6
- 235000019418 amylase Nutrition 0.000 description 6
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 6
- 230000001954 sterilising effect Effects 0.000 description 6
- 229910001629 magnesium chloride Inorganic materials 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 238000000197 pyrolysis Methods 0.000 description 4
- 238000011218 seed culture Methods 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 3
- 241000556413 Caldicellulosiruptor owensensis Species 0.000 description 3
- 229910021578 Iron(III) chloride Inorganic materials 0.000 description 3
- 239000007836 KH2PO4 Substances 0.000 description 3
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 3
- 239000001110 calcium chloride Substances 0.000 description 3
- 229910001628 calcium chloride Inorganic materials 0.000 description 3
- 229960002433 cysteine Drugs 0.000 description 3
- 230000001461 cytolytic effect Effects 0.000 description 3
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 3
- 230000002255 enzymatic effect Effects 0.000 description 3
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 3
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 3
- 229910052603 melanterite Inorganic materials 0.000 description 3
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 241000589516 Pseudomonas Species 0.000 description 2
- 235000013339 cereals Nutrition 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 1
- 241001429558 Caldicellulosiruptor bescii Species 0.000 description 1
- 241000987689 Caldicellulosiruptor hydrothermalis Species 0.000 description 1
- 241000511681 Caldicellulosiruptor kristjanssonii Species 0.000 description 1
- 241000152015 Caldicellulosiruptor obsidiansis Species 0.000 description 1
- 241000178335 Caldicellulosiruptor saccharolyticus Species 0.000 description 1
- 241001560086 Pachyrhizus Species 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 108010089934 carbohydrase Proteins 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 238000011090 industrial biotechnology method and process Methods 0.000 description 1
- 239000010985 leather Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000001814 pectin Substances 0.000 description 1
- 229920001277 pectin Polymers 0.000 description 1
- 235000010987 pectin Nutrition 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 241001148471 unidentified anaerobic bacterium Species 0.000 description 1
- 229920001221 xylan Polymers 0.000 description 1
- 150000004823 xylans Chemical class 0.000 description 1
- 150000008494 α-glucosides Chemical class 0.000 description 1
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
Sweet potato waste is that sweet potato powder factory extracts the residue after starch from sweet potato, accounts for about the 45%-60% of fresh weight.The main component of sweet potato waste is starch, cellulose and a small amount of protein.Starch-containing 36-58% in dry sweet potato waste, cellulose 20-30% can produce the glucose for accounting for dry weight 80% or so if it is digested completely, be the good sources of carbon of the fermenting and producing of large fermented product such as ethyl alcohol, butanol etc..Sweet potato waste is most of not utilized in addition to small part is used to produce dietary fiber, feed and solid fuel at present, or even becomes pollution sources.The present invention provides a kind of method using high temperature enzyme rapid enzymolysis sweet potato waste, by hot fermentation generate crude enzyme liquid and sweet potato waste be mixed in a certain proportion after 60-80 DEG C enzymatic hydrolysis 10-20 hours, starch and cellulase hydrolysis rate in sweet potato waste reach 90%-100%.
Description
Technical field
The invention belongs to field of biotechnology, in particular to a kind of sweet potato waste rapid enzymolysis at monosaccharide method.
Background technique
Sweet potato is also known as sweet potato, sweet potato, red tongue fur, sweet potato, pachyrhizus, cultivates extensively in global tropical, subtropical zone
Area.Sweet potato is annual herb plant, and under ground portion is round, ellipse or fusiform root tuber, the shape of root tuber, color of the leather and
Yellowish pink is different because of kind or soil difference.Sweet potato is important one of the cereal crops in China, is almost all distributed throughout the country,
Major production areas is the ground such as Hunan, Hubei, Jiangxi, Anhui and Shandong.According to statistics, China sweet potato total output accounts for the world up to 100,000,000 tons
80% or so of sweet potato total output.Sweet potato waste be sweet potato powder factory production sweet potato powder after and starch from sweet potato factory extract starch from sweet potato after
Residue accounts for about the 45%-60% of fresh weight.The main component of sweet potato waste is starch, cellulose and a small amount of protein.Wherein starch and
The content of cellulose accounts for the 36-58% and 20-30% of sweet potato waste dry weight respectively, is degraded into the monosaccharide based on glucose,
It is the very good material of fermentation industry.Monosaccharide is degraded into applied to fermentation industry using sweet potato waste, can be relieved fermentation industry to grain
Dependence, while be expected reduce production cost, promote fermentation industry sustainable health development.
However sweet potato waste is not fully used at present, except small part is for producing dietary fiber, feed and solid
It is most of to become pollutant outside fuel.To find out its cause, mainly without economic, quick biodegrading process.Chemically divide in structure
Analysis, although starch and cellulose are all to have glucose polymerisation to form, they are two different high polymers.Starch is by α glucosides
Key link, and cellulose is formed by β-Isosorbide-5-Nitrae glycosidic bond links, is formed simultaneously crystalline texture.Therefore, they digest the enzyme being related to
Type and enzymatic hydrolysis condition are all different, and amylorrhexis is mainly completed by amylase and carbohydrase, and the enzymatic hydrolysis of cellulose is by fiber
Plain enzyme is completed.The enzymatic hydrolysis condition difference of current commercialized amylase and cellulase is very big, is difficult that starch and cellulose is same
When degrade.
Thermophilic Bacterium is a kind of microorganism that can be grown at relatively high temperatures, the active high, heat of generated enzyme
The advantages that stability is good has important application value in industrial biotechnology field.Wherein it is pyrolyzed cellulose Pseudomonas
It (Caldicellulosiruptor) is the most heat-resisting a kind of biomass economy anaerobic bacteria found so far, so far, pyrolysis is fine
Tieing up plain Pseudomonas has had 8 kinds of (C.saccharolyticus, C.bescii, C.obsidiansis
C.hydrothermalis, C.kristjanssonii, C.kronotskyensis, C.lactoaceticus, and
C.owensensis it) is successively found, can be utilized under high temperature (70-78 DEG C) on Europe, North America, Asia and Australia and other places
The high temperature enzyme of the growth such as starch, cellulose, pectin, xylan, secretion has considerable prospects for commercial application.This hair
It is bright using sweet potato waste as inducer, individually using pyrolysis CELLULOLYTIC BACTERIUM Caldicellulosiruptor belong to bacterial strain, such as
Caldicellulosiruptor owensensis (DSM13100), Caldicellulosiruptor kronotskyensis
(DSM19802), Caldicellulosiruptor lactoaceticus (DSM9545) etc. is (purchased from German microbial preservation
Heart DSMZ) fermentation, the crude enzyme liquid for existing simultaneously amylase and cellulase is obtained, crude enzyme liquid is directly used in the quick enzyme of sweet potato waste
Solution.Due in crude enzyme liquid amylase and cellulase be by same plant height temperature bacterium generate, enzymatic hydrolysis condition is very close,
Preferable hydrolysis result can be reached under mutually synthermal and pH condition, it can be quickly by starch and cellulose degradation at monosaccharide.It can
To find out that the present invention is easily operated, there is huge industrial application value.
Summary of the invention
[purpose of the present invention]
It is digested the purpose of the present invention is sweet potato waste is efficient, economical, quickly generates fermentable sugars, provided for fermentation industry honest and clean
Valence raw material, realizes the higher value application of sweet potato waste, while reducing the environmental pollution of sweet potato powder production process.
[technical solution]
For sweet potato waste rapid enzymolysis provided by the invention at the method for monosaccharide, technical solution is as follows:
1) preparation of crude enzyme liquid: pyrolysis CELLULOLYTIC BACTERIUM grows 12-16h at 70-80 DEG C in seed culture medium, obtains seed
Seed liquor is inoculated into according to 0.4%-1% containing using sweet potato waste as the fermentation medium of inducer, in 60-80 DEG C of liquid by liquid
Fermentation liquid is centrifuged 5-8 minutes under the conditions of 4000-5000 revs/min after fermentation, obtains supernatant by anaerobic fermentation 10-16h
Liquid is crude enzyme liquid;
2) enzymatic hydrolysis of sweet potato waste: obtained crude enzyme liquid is mixed according to the amount of every gram of dry sweet potato slag of 0.1-1ml with sweet potato waste,
Water supplement be prepared into solid content be 10-20% mixture, 10-20h is digested at 60-80 DEG C, the starch in sweet potato waste and
The enzymatic hydrolyzation of cellulose reaches 90%-100%.
[technical characterstic]
The present invention has the characteristics that amylase and cellulase abundant using pyrolysis CELLULOLYTIC BACTERIUM, by being lured with sweet potato waste
Amylase and cellulase that fermentation generates high activity are led, and they can play a role under identical enzymatic hydrolysis condition, with fermentation
Obtained crude enzyme liquid just by the starch and cellulose in sweet potato waste while can degrade, and requirement of the present invention to process equipment be not severe
It carves, is easy to industrial applications.
Specific embodiment
Embodiment 1
Strain Caldicellulosiruptor owensensis (DSM13100) is in seed culture medium (1.3g (NH4)2SO4,1.5gKH2PO4,2.9g K2HPO4·3H2O,0.2g MgCl2·6H2O,0.075g CaCl2·2H2O,1.250mg
FeSO4·7H2O, 2g yeast powder, 0.5g Cysteine-HClH2O, 6g glucose add water constant volume to 1L, by pH tune before sterilizing
To 75 DEG C of culture 12h in 7.2), fermentation medium (NH4Cl0.90g, NaCl0.90g, MgCl are inoculated into according to 1% amount2·
6H2O0.40g, KH2PO40.75g, K2HPO41.50g, Trypticase2.00g, yeast powder 1.00g, FeCl3·
6H2O2.50mg, sweet potato waste 1.00g, Cysteine-HCl x H27.2) pH is transferred to before sterilizing by O0.75g with water constant volume at 1L
In, 70 DEG C of culture 15h are centrifugated thallus, supernatant are taken to mix according to the amount of every gram of dry sweet potato slag of 0.3ml with sweet potato waste, mend
Add water to be prepared into the mixture that solid content is 10%, 10h is digested at 80 DEG C, the enzymatic hydrolysis of starch and cellulose in sweet potato waste
Rate respectively reaches 98% and 95%.
Embodiment 2
Strain Caldicellulosiruptor kronotskyensis (DSM19802) is in seed culture medium (1.3g
(NH4)2SO4,1.5gKH2PO4,2.9g K2HPO4·3H2O,0.2g MgCl2·6H2O,0.075g CaCl2·2H2O,
1.250mg FeSO4·7H2O, 2g yeast powder, 0.5g Cysteine-HClH2O, 6g glucose add water constant volume to 1L, sterilizing
It is preceding that pH is transferred in 7.2), 73 DEG C of culture 15h, according to 0.5% amount be inoculated into fermentation medium (NH4Cl0.90g,
NaCl0.90g, MgCl2·6H2O0.40g, KH2PO40.75g, K2HPO41.50g, Trypticase2.00g, yeast powder 1.00g,
FeCl3·6H2O2.50mg, sweet potato waste 1.00g, Cysteine-HCl x H2O0.75g, with water constant volume at 1L, by pH before sterilizing
It is transferred in 7.2), 75 DEG C of culture 15h, is centrifugated thallus, takes supernatant according to the amount and sweet potato waste of every gram of dry sweet potato slag of 0.5ml
Mixing, water supplement are prepared into the mixture that solid content is 20%, 16h are digested at 70 DEG C, starch and fiber in sweet potato waste
The enzymatic hydrolyzation of element respectively reaches 99% and 96%.
Embodiment 3
Strain Caldicellulosiruptor lactoaceticus (DSM9545) is in seed culture medium (1.3g
(NH4)2SO4,1.5gKH2PO4,2.9g K2HPO4·3H2O,0.2g MgCl2·6H2O,0.075g CaCl2·2H2O,
1.250mg FeSO4·7H2O, 2g yeast powder, 0.5g Cysteine-HClH2O, 6g glucose add water constant volume to 1L, sterilizing
It is preceding that pH is transferred in 7.2), 77 DEG C of culture 12h, according to 0.8% amount be inoculated into fermentation medium (NH4Cl0.90g,
NaCl0.90g, MgCl2·6H2O0.40g, KH2PO40.75g, K2HPO41.50g, Trypticase2.00g, yeast powder 1.00g,
FeCl3·6H2O2.50mg, sweet potato waste 1.00g, Cysteine-HCl x H2O0.75g, with water constant volume at 1L, by pH before sterilizing
It is transferred in 7.2), 78 DEG C of culture 15h, is centrifugated thallus, take supernatant mixed according to the amount and sweet potato waste of every gram of dry sweet potato slag of 1ml
It closes, water supplement is prepared into the mixture that solid content is 15%, 14h is digested at 75 DEG C, starch and cellulose in sweet potato waste
Enzymatic hydrolyzation respectively reach 100% and 90%.
Claims (1)
1. a kind of sweet potato waste rapid enzymolysis includes the following steps: at the method for monosaccharide
(1) high temperature bacterium is using sweet potato waste as inducer, and in 70-75 DEG C of liquid anaerobic fermentation 10-16h, the high temperature bacterium bag is included
Caldicellulosiruptor.owensensis (DSM 13100) and Caldicellulosiruptor
kronotskyensis(DSM 19802),Caldicellulosiruptor lactoaceticus(DSM 9545);
(2) it is centrifugated thallus after fermenting, obtains crude enzyme liquid;
(3) crude enzyme liquid that step (2) obtains is mixed according to the amount of every gram of dry sweet potato slag of 0.1-1ml with sweet potato waste, water supplement preparation
The mixture for being 10-20% at solid content, digests 10-20h, the enzyme of starch and cellulose in sweet potato waste at 60-80 DEG C
Solution rate reaches 90%-100%.
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101851650A (en) * | 2010-04-19 | 2010-10-06 | 中国科学院青岛生物能源与过程研究所 | Method for saccharifying cellulose raw material |
CN102286600A (en) * | 2011-08-08 | 2011-12-21 | 华南理工大学 | Method for simultaneously producing ethanol and hydrogen by using cassava residue through fermentation |
CN102559811A (en) * | 2012-03-15 | 2012-07-11 | 吴允山 | Method of preparing sugar by utilizing sweet potato residues |
CN102618602A (en) * | 2012-03-30 | 2012-08-01 | 吴允山 | Process for preparing sugar by performing enzymatic hydrolysis on sweet potato residues |
-
2014
- 2014-05-12 CN CN201410198435.1A patent/CN105087716B/en not_active Expired - Fee Related
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101851650A (en) * | 2010-04-19 | 2010-10-06 | 中国科学院青岛生物能源与过程研究所 | Method for saccharifying cellulose raw material |
CN102286600A (en) * | 2011-08-08 | 2011-12-21 | 华南理工大学 | Method for simultaneously producing ethanol and hydrogen by using cassava residue through fermentation |
CN102559811A (en) * | 2012-03-15 | 2012-07-11 | 吴允山 | Method of preparing sugar by utilizing sweet potato residues |
CN102618602A (en) * | 2012-03-30 | 2012-08-01 | 吴允山 | Process for preparing sugar by performing enzymatic hydrolysis on sweet potato residues |
Non-Patent Citations (2)
Title |
---|
《Complete Genome Sequences for the Anaerobic, Extremely Thermophilic》;Sara E.等;《JOURNAL OF BACTERIOLOGY》;20110107;第193卷(第6期);全文 * |
《极端嗜热厌氧菌Caldicellulosiruptor木质纤维素降解研究》;孟冬冬等;《生物加工过程》;20040131;第12卷(第1期);第38页2.1节第一段;参见第41页第一段以及表1 * |
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