CN100448987C - Alpha-acetyl lactate dehydrogenase dry powder and its preparation process - Google Patents
Alpha-acetyl lactate dehydrogenase dry powder and its preparation process Download PDFInfo
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- CN100448987C CN100448987C CNB2004100609872A CN200410060987A CN100448987C CN 100448987 C CN100448987 C CN 100448987C CN B2004100609872 A CNB2004100609872 A CN B2004100609872A CN 200410060987 A CN200410060987 A CN 200410060987A CN 100448987 C CN100448987 C CN 100448987C
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- alpha
- spray
- enzyme
- acetolactate decarboxylase
- dry powder
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- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 239000000843 powder Substances 0.000 title claims description 19
- 102000003855 L-lactate dehydrogenase Human genes 0.000 title 1
- 108700023483 L-lactate dehydrogenases Proteins 0.000 title 1
- 108090000790 Enzymes Proteins 0.000 claims abstract description 50
- 102000004190 Enzymes Human genes 0.000 claims abstract description 50
- 108010084631 acetolactate decarboxylase Proteins 0.000 claims abstract description 19
- 239000007788 liquid Substances 0.000 claims abstract description 19
- 235000010413 sodium alginate Nutrition 0.000 claims abstract description 7
- 229920001285 xanthan gum Polymers 0.000 claims abstract description 7
- 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 claims abstract description 6
- 239000000661 sodium alginate Substances 0.000 claims abstract description 6
- 229940005550 sodium alginate Drugs 0.000 claims abstract description 6
- 229920001353 Dextrin Polymers 0.000 claims abstract description 5
- 239000000463 material Substances 0.000 claims description 18
- 230000001681 protective effect Effects 0.000 claims description 18
- 230000000694 effects Effects 0.000 claims description 15
- 239000007921 spray Substances 0.000 claims description 13
- 229920002774 Maltodextrin Polymers 0.000 claims description 6
- 239000012141 concentrate Substances 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
- 235000010493 xanthan gum Nutrition 0.000 claims description 6
- 239000000230 xanthan gum Substances 0.000 claims description 6
- 229940082509 xanthan gum Drugs 0.000 claims description 6
- 239000000047 product Substances 0.000 claims description 5
- WHGYBXFWUBPSRW-UHFFFAOYSA-N Cycloheptaamylose Natural products O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO WHGYBXFWUBPSRW-UHFFFAOYSA-N 0.000 claims description 4
- 239000004375 Dextrin Substances 0.000 claims description 4
- 239000005913 Maltodextrin Substances 0.000 claims description 4
- WHGYBXFWUBPSRW-FOUAGVGXSA-N beta-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO WHGYBXFWUBPSRW-FOUAGVGXSA-N 0.000 claims description 4
- 235000019425 dextrin Nutrition 0.000 claims description 4
- 229940035034 maltodextrin Drugs 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 2
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims description 2
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims description 2
- 230000001580 bacterial effect Effects 0.000 claims description 2
- 238000007796 conventional method Methods 0.000 claims description 2
- 239000007787 solid Substances 0.000 claims description 2
- 238000003756 stirring Methods 0.000 claims description 2
- 239000006228 supernatant Substances 0.000 claims description 2
- 238000001694 spray drying Methods 0.000 abstract 5
- 239000003223 protective agent Substances 0.000 abstract 2
- DPVHGFAJLZWDOC-PVXXTIHASA-N (2r,3s,4s,5r,6r)-2-(hydroxymethyl)-6-[(2r,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxane-3,4,5-triol;dihydrate Chemical compound O.O.O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 DPVHGFAJLZWDOC-PVXXTIHASA-N 0.000 abstract 1
- GJCOSYZMQJWQCA-UHFFFAOYSA-N 9H-xanthene Chemical compound C1=CC=C2CC3=CC=CC=C3OC2=C1 GJCOSYZMQJWQCA-UHFFFAOYSA-N 0.000 abstract 1
- FYGDTMLNYKFZSV-MRCIVHHJSA-N dextrin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)OC1O[C@@H]1[C@@H](CO)OC(O[C@@H]2[C@H](O[C@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-MRCIVHHJSA-N 0.000 abstract 1
- 235000013373 food additive Nutrition 0.000 abstract 1
- 239000002778 food additive Substances 0.000 abstract 1
- 229940074409 trehalose dihydrate Drugs 0.000 abstract 1
- 235000013405 beer Nutrition 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 5
- 238000012856 packing Methods 0.000 description 4
- 238000003860 storage Methods 0.000 description 4
- 150000003625 trehaloses Chemical class 0.000 description 4
- ROWKJAVDOGWPAT-UHFFFAOYSA-N Acetoin Chemical compound CC(O)C(C)=O ROWKJAVDOGWPAT-UHFFFAOYSA-N 0.000 description 2
- 241000194108 Bacillus licheniformis Species 0.000 description 2
- 241000193764 Brevibacillus brevis Species 0.000 description 2
- 244000057717 Streptococcus lactis Species 0.000 description 2
- 235000014897 Streptococcus lactis Nutrition 0.000 description 2
- 238000006114 decarboxylation reaction Methods 0.000 description 2
- 239000008297 liquid dosage form Substances 0.000 description 2
- 241000589220 Acetobacter Species 0.000 description 1
- 241000589212 Acetobacter pasteurianus Species 0.000 description 1
- 108010002945 Acetoin dehydrogenase Proteins 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 101000836529 Brevibacillus brevis Alpha-acetolactate decarboxylase Proteins 0.000 description 1
- 102100027269 Fructose-bisphosphate aldolase C Human genes 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 239000012752 auxiliary agent Substances 0.000 description 1
- OWBTYPJTUOEWEK-UHFFFAOYSA-N butane-2,3-diol Chemical class CC(O)C(C)O OWBTYPJTUOEWEK-UHFFFAOYSA-N 0.000 description 1
- 238000005138 cryopreservation Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- GFAZHVHNLUBROE-UHFFFAOYSA-N hydroxymethyl propionaldehyde Natural products CCC(=O)CO GFAZHVHNLUBROE-UHFFFAOYSA-N 0.000 description 1
- 239000013067 intermediate product Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
Abstract
The present invention discloses spray-drying alpha-acetolactate decarboxylase and a preparation method thereof, the spray-drying alpha-acetolactate decarboxylase belongs to a food additive biological product of an enzyme preparation and comprises liquid alpha-acetolactate decarboxylase and a spray-drying protective agent, wherein the spray-drying protective agent is composed of trehalose dihydrate, xanthan, sodium alginate, dextrine, etc., and is prepared by a corresponding spray-drying technology. The enzyme has high stability and can be effectively saved for 1 to 2 years at dry places at normal temperature.
Description
Technical field
The food that the present invention relates to a kind of zymin adds biological products, is specifically related to a kind of alpha-acetolactate decarboxylase dry powder and preparation method thereof.
Background technology
(ALDC EC4.1.1.5) has obtained the widespread use of beer production producer to alpha-acetolactate decarboxylase as the auxiliary agent of beer slaking.In the brewage process, α-acetylactis forms di-acetyl through non-oxydasis decarboxylation, when the content of di-acetyl surpasses 0.15mg/kg, has a strong impact on the taste of beer.Traditional method of removing di-acetyl is a low-temperature storage beer, and the diacetyl reductase that allows yeast itself produce makes di-acetyl finally be reduced into 2,3 butyleneglycols, and this process generally needed for 3～6 weeks the beer maturing process, and what have reaches 3 months.This enzyme is applied in the beer production, and it can change into acetoin with α-acetylactis direct oxidation decarboxylation, without this intermediate product of di-acetyl, thereby can make fermentation period shorten 3～4 times, and usage ratio of equipment improves 40%.Not only can shorten the production cycle of beer greatly, and make finished beer not produce di-acetyl rise phenomenon in storage process, constant product quality reduces production costs.At present, this enzyme is liquid dosage form, and stability is not good enough, needs the transportation of cryopreservation and van cooler, and the stability of enzyme is manufacturer and uses the problem that producer all very pays close attention to.
By retrieval, find some and extract and produce the scientific and technical literature of alpha-acetolactate decarboxylase, but do not see the research report that alpha-acetolactate decarboxylase dry powder is arranged from bacterial classification.The alpha-acetolactate decarboxylase of manufacturer's production both at home and abroad is liquid dosage form at present, and thermostability is relatively poor, is difficult for storage and transport.
Summary of the invention
The object of the present invention is to provide a kind of Heat stability is good, be convenient to alpha-acetolactate decarboxylase dry powder of storage and transport and preparation method thereof.
The following surface technology scheme of the present invention realizes:
The preparation method of alpha-acetolactate decarboxylase dry powder comprises that earlier enzyme being produced bacterium Lactococcus lactis (Lactococcus lactis) or subtilis (Bacillus subtilis) or weak oxide acetobacter (Acetobacter pasteurianus) or bacillus brevis (Bacillus brevis) or Bacillus licheniformis (Bacillus licheniformis) inoculates according to a conventional method and ferment; obtain behind the leavened prod centrifugal; getting supernatant concentration is 10-30% to containing amount of solid; obtain concentrating enzyme liquid; in concentrated enzyme liquid, add protective material then; add protectant amount for concentrating the 1-30% of enzyme liquid total amount; stir; protective material is dissolved in fully concentrates in the enzyme liquid; through behind the homogeneous; in inlet temperature is 150-2002; temperature of outgoing air is 60-90 ℃; cyclone separating tower pressure reduction is 0.7 * 100KPa; feed rate be spray on the Highspeedcentrifugingandsprayingdrier of 30-50 ml/min dried; promptly get the dried enzyme powder of flaxen spray, be packaged to be product again.This product is contained in and places dry place in the plastics bag, can effectively preserve 1-2 under the condition of normal temperature (25 ℃).
Above-described protectant composition and parts by weight are as follows: 0.5～20 part of trehalose, and 0.01～0.5 part of xanthan gum, 1～20 part in dextrin, 0.2～2.0 part of sodium alginate,
Dextrin wherein can be cycloheptaamylose or maltodextrin, or cycloheptaamylose and maltodextrin are with arbitrary proportion blended mixture.
The alpha-acetolactate decarboxylase dry powder for preparing with the present invention should detect its enzyme activity and water content, technical indicator: enzyme activity 〉=10000u/g, moisture content≤5%.
The active character that alpha-acetolactate decarboxylase adding protective material of the present invention is made behind the dry powder does not change; identical with liquid enzyme; but the transportation and use more convenient; as long as add distilled water or pure water dilution, greatly facilitate the user like this before using, the past is because alpha-acetolactate decarboxylase is liquid; must be kept in 0-10 ℃ the freezer; and adopt the low temperature transportation, if run into the enzyme activity reduction in transit of weather temperature height, influenced the quality of enzyme.
Embodiment
Embodiment one
1. by following set of dispense system protective material enzyme liquor: (w/v)
Concentrate 100 milliliters of enzyme liquid
Protective material 10 grams, wherein:
5 parts of trehaloses
0.08 part of xanthan gum
25 parts of maltodextrins
0.42 part of sodium alginate
2. each component of protective material is joined in the enzyme liquid, it is back standby to be stirred to dissolving fully.
3. be 30～40MPa in operating pressure, working flow be carry out on 1 liter/minute the high pressure homogenizer after the second homogenate standby.
4. be 150 ℃ in inlet temperature, temperature of outgoing air is 90 ℃, and cyclone separating tower pressure reduction is 0.7 * 100KPa, and feed rate is to spray driedly on the Highspeedcentrifugingandsprayingdrier of 30 ml/min, promptly gets the dried enzyme powder of flaxen spray.
5. detect its enzyme activity and water content, enzyme activity is answered 〉=10000u/g, and moisture content answers≤5%.
6. packing places dry place, and normal temperature is preserved down.
Embodiment two
1. by following set of dispense system protective material enzyme liquor: (w/v)
Concentrate 100 milliliters of enzyme liquid
Protective material 1 gram, wherein:
20 parts of trehaloses
0.12 part of xanthan gum
10 parts of cycloheptaamyloses
2.0 parts of sodium alginates
2. each component of protective material is joined in the enzyme liquid, it is back standby to be stirred to dissolving fully.
3. be 30～40MPa in operating pressure, working flow be carry out on 1 liter/minute the high pressure homogenizer behind homogeneous standby.
4. be 160 ℃ in inlet temperature, temperature of outgoing air is 80 ℃, and cyclone separating tower pressure reduction is 0.7 * 100KPa, and feed rate is to spray driedly on the Highspeedcentrifugingandsprayingdrier of 40 ml/min, promptly gets the dried enzyme powder of flaxen spray.
5. detect its enzyme activity and water content, enzyme activity is answered 〉=10000u/g, and moisture content answers≤5%.
6. packing places dry place, and normal temperature is preserved down.
Embodiment three
1. by following set of dispense system protective material enzyme liquor: (w/v)
Concentrate 100 milliliters of enzyme liquid
Protective material 30 grams, wherein:
15 parts of trehaloses
0.15 part of xanthan gum
15 parts of maltodextrins
0.35 part of sodium alginate
2. each component of protective material is joined in the enzyme liquid, it is back standby to be stirred to dissolving fully.
3. be 30～40MPa in operating pressure, working flow be carry out on 1 liter/minute the high pressure homogenizer after the second homogenate standby.
4. be 170 ℃ in inlet temperature, temperature of outgoing air is 80 ℃, and cyclone separating tower pressure reduction is 0.7 * 100KPa, and feed rate is to spray driedly on the Highspeedcentrifugingandsprayingdrier of 50 ml/min, promptly gets the dried enzyme powder of flaxen spray.
5. detect its enzyme activity power and water content, enzyme activity power is answered 〉=10000u/g, and moisture content answers≤5%.
6. packing places dry place, and normal temperature is preserved down.
Embodiment four
1. by following set of dispense system protective material enzyme liquor: (w/v)
Concentrate 100 milliliters of enzyme liquid
Protective material 25 grams, wherein:
25 parts of trehaloses
0.05 part of xanthan gum
5 parts of cycloheptaamyloses
0.45 part of sodium alginate
2. each component of protective material is joined in the enzyme liquid, it is back standby to be stirred to dissolving fully.
3. be 30～40MPa in operating pressure, working flow be carry out on 1 liter/minute the high pressure homogenizer after the second homogenate standby.
4. be 200 ℃ in inlet temperature, temperature of outgoing air is 70 ℃, and cyclone separating tower pressure reduction is 0.7 * 100KPa, and feed rate is to spray driedly on the Highspeedcentrifugingandsprayingdrier of 50 ml/min, promptly gets the dried enzyme powder of flaxen spray.
5. detect its enzyme activity and water content, enzyme activity is answered 〉=10000u/g, and moisture content answers≤5%.
6. packing places dry place, and normal temperature is preserved down.
The foregoing description has provided preferred operations condition of the present invention, but technology contents of the present invention is not subjected to the limitation of embodiment.It is as follows to spray dried enzyme powder acceleration thermal stability test result:
50 ℃ 7 days, front and back enzyme activity rate of descent≤1%.
50 ℃ 14 days, front and back enzyme activity rate of descent≤2%.
The dry place of normal temperature (25 ℃) 24 months, enzyme activity rate of descent≤5%.
Claims (4)
1, a kind of preparation method of alpha-acetolactate decarboxylase dry powder, comprise the process that bacterial classification is inoculated according to a conventional method and fermented, it is characterized in that: after obtaining leavened prod, earlier centrifugal, getting supernatant concentration is 10-30% to containing amount of solid, obtain concentrating enzyme liquid, in concentrated enzyme liquid, add protective material then, add protectant amount for concentrating the 1-30% of enzyme liquid total amount, stir, protective material is dissolved in fully concentrates in the enzyme liquid, through behind the homogeneous, on spray-drier, spray driedly, obtain the dried enzyme powder of flaxen spray;
Described protectant composition and parts by weight are as follows: 0.5～20 part of trehalose, 0.01～0.5 part of xanthan gum, 1～20 part in dextrin, 0.2～2.0 part of sodium alginate.
2, the preparation method of alpha-acetolactate decarboxylase dry powder according to claim 1 is characterized in that: the dextrin in the protective material is cycloheptaamylose or maltodextrin, or cycloheptaamylose and maltodextrin are with arbitrary proportion blended mixture.
3, the preparation method of alpha-acetolactate decarboxylase dry powder according to claim 1, it is characterized in that: spray-dired inlet temperature is 150-200 ℃, temperature of outgoing air is 60-90 ℃, and cyclone separating tower pressure reduction is 0.7 * 100KPa, and feed rate is the 30-50 ml/min.
4, the preparation method of alpha-acetolactate decarboxylase dry powder according to claim 1 is characterized in that: in the alpha-acetolactate decarboxylase power-product for preparing, and its enzyme activity 〉=10000u/g, moisture content≤5%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100609872A CN100448987C (en) | 2004-10-18 | 2004-10-18 | Alpha-acetyl lactate dehydrogenase dry powder and its preparation process |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100609872A CN100448987C (en) | 2004-10-18 | 2004-10-18 | Alpha-acetyl lactate dehydrogenase dry powder and its preparation process |
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| Publication Number | Publication Date |
|---|---|
| CN1624121A CN1624121A (en) | 2005-06-08 |
| CN100448987C true CN100448987C (en) | 2009-01-07 |
Family
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| Application Number | Title | Priority Date | Filing Date |
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| CNB2004100609872A Expired - Fee Related CN100448987C (en) | 2004-10-18 | 2004-10-18 | Alpha-acetyl lactate dehydrogenase dry powder and its preparation process |
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Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016191169A1 (en) * | 2015-05-22 | 2016-12-01 | Dupont Nutrition Biosciences Aps | Acetolactate decarboxylase |
| CN110669752A (en) * | 2019-11-12 | 2020-01-10 | 河北省微生物研究所 | Preparation method of hexokinase spray-dried powder |
-
2004
- 2004-10-18 CN CNB2004100609872A patent/CN100448987C/en not_active Expired - Fee Related
Non-Patent Citations (6)
| Title |
|---|
| A-乙酰乳酸脱羧酶的3种固定化方法的比较. 钱斯日古楞.大连轻工业学院学报，第21卷第4期. 2002 |
| A-乙酰乳酸脱羧酶的3种固定化方法的比较. 钱斯日古楞.大连轻工业学院学报，第21卷第4期. 2002 * |
| a-乙酰乳酸脱羧酶的研究进展及其应用前景. 张博润等.微生物学通报，第24卷第4期. 1997 |
| a-乙酰乳酸脱羧酶的研究进展及其应用前景. 张博润等.微生物学通报，第24卷第4期. 1997 * |
| 磁性淀粉微球固定化乙酰乳酸脱羧酶及应用. 邱广亮等.广州化工，第28卷第4期. 2000 |
| 磁性淀粉微球固定化乙酰乳酸脱羧酶及应用. 邱广亮等.广州化工，第28卷第4期. 2000 * |
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| CN1624121A (en) | 2005-06-08 |
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Address after: No. 60, orchard Avenue, hi tech Industrial Development Zone, the Guangxi Zhuang Autonomous Region, Nanning Patentee after: Bangerke Biological Technology Co., Ltd., Nanning Address before: The Guangxi Zhuang Autonomous Region, Nanning hi tech Industrial Development Zone, Science Park Road, No. five East Fourth Avenue Patentee before: Bangerke Biological Technology Co., Ltd., Nanning |
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Granted publication date: 20090107 Termination date: 20191018 |