CN102286419A - Enterobacter SYA2 and method for preparing lactase with same - Google Patents
Enterobacter SYA2 and method for preparing lactase with same Download PDFInfo
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Abstract
The invention discloses an enterobacter SYA2 and a method for preparing lactase with the same. The bacterial strain is an enterobacter, and is screened from yak milk yoghurt collected from a pasturing area in Gannan prefecture of Gansu province by adopting screening culture medium containing X-gal. The method for preparing the lactase includes the following steps: the enterobacter as the strain is adopted to prepare fermentation broth, the obtained fermentation broth undergoes high-speed refrigerated centrifugation, ammonium sulfate fractionation precipitation, DEAE-SephadexA-25 ion-exchange column chromatography, membrane filtration and concentration and freeze drying, and thereby a finished lactase product, which can be used in the processing of dairy products, is obtained.
Description
Technical field
The invention belongs to technical field of bioengineering, a kind of specifically enterobacteria and prepare the method for Sumylact L with it.
Background technology
Sumylact L (Lactase) formal name used at school is β-D-galactoside galactohydrolase (β-D-galactoside galactohydrolase, E.C.3.2.1.23), have another name called beta-galactosidase enzymes (β-galactosidase), be the enzyme of a class catalysis β-D-semi-lactosi glycosidic bond generation hydrolysis, it can hydrolyze lactose into semi-lactosi and glucose.Sumylact L is widely distributed in animal, plant and microorganism.
So far, discover that the microorganism that can produce Sumylact L has a lot, common mould comprise aspergillus niger (Aspergillus niger), aspergillus oryzae (
Aspergillus oryzae), Aspergillus candidus (
Aspergillus candidus), Mucor pusillus (
Mucor pusillus) etc.; Yeast comprise Kluyveromyces fragilis (
Kluyveromyces fragilis), Kluyveromyces lactis (
Kluyveromyces lactis), candida tropicalis (
Candida tropicalis) etc.; Bacterium comprise intestinal bacteria (
Escherichia Coli), enterobacter agglomerans (
Enterobacter agglomerans), bacillus megaterium (
Bacillus megaterium), subtilis (
Bacillus subtilis), Bacillus circulans (
Bacillus Circulans), bacstearothermophilus (
Bacillus stearothermophilus), lactobacillus bulgaricus (
Lactobacillus bulgaricus), bifidus bacillus (
Bifidobacterium), aquatic draw the engler bacterium (
Rahnella aquatilis), thermophilus streptococcus (
Streptococcus thermophilus), Arthrobacter (
Arthrobacter aurescens), Flavobacterium (
Flavobacterium) etc.The Sumylact L character and the purposes difference in different microorganisms source: mould acts on optimal pH slant acidity (pH2.5～5.0), is mainly used in the hydrolysis of acid whey and cheese; Yeast institute galactopoiesis carbohydrase optimal pH is neutrality (pH6.0～7.0) closely, is suitable for the processing treatment of cow's milk and whey; Enzyme that bacterium produced and yeast optimal pH near and have a higher thermotolerance.
The research of bacterium Sumylact L still is in the starting stage both at home and abroad at present.
Summary of the invention
The purpose of this invention is to provide a kind of enterobacteria SYA2.
Another object of the present invention provides a kind of method for preparing Sumylact L with enterobacteria.
Enterobacteria SYA2 among the present invention gathers from pastoral area, state, Gannan, Gansu Province to screen the yak yoghourt and obtains, classification called after enterobacteria (
Enterobacter sp) SYA2, on 09 13rd, 2011 in China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC) preservation, preserving number CGMCC No:5251.
The screening method of enterobacteria SYA2 of the present invention is to gather pastoral area, state, Gannan, Gansu Province to gather yak yoghourt sample, coat on the separation screening substratum that contains X-gal, observe colony colour and become blue degree and carry out preliminary screening, and lactase activity size and obtaining relatively.The pastoral area pedotheque is selected well-grown, color to become the blue big bacterium colony of degree and is carried out the shake flask fermentation cultivation through the separation screening culture medium culturing, by comparing the lactase activity size, finally determines bacterial strain.
The separation screening substratum that contains X-gal is formed: lactose 20 g/L, yeast extract paste 10 g/L, peptone 10g/L, X-gal 0.04 g/L, agar 20 g/L, pH 7.0.
A standard enzyme unit alive (U) is defined as: discharge the required enzyme amount of 1 μ moL ONP 37 ℃ of per minute hydrolysis.Get the crude enzyme liquid after the 0.5mL dilution, at 37 ℃ of following water-bath 5min, add be preheated to 37 ℃ contain 5mmol/L ONPG phosphate buffered saline buffer (pH6.5) 1.5mL, 37 ℃ of following water-bath 10min add 3.0mL 0.5mol/LNa then immediately
2CO
3Termination reaction is measured the OD value in the 420nm place, measures 3 times and averages.
In the formula, X is the Sumylact L vigor; C is an ONP concentration corresponding on the typical curve; N is the extension rate of enzyme liquid.
Enterobacteria SYA2 CGMCC No:5251 of the present invention has the ability of galactopoiesis carbohydrase.
The present invention is following steps with the method that enterobacteria SYA2 prepares Sumylact L:
A, seed liquor preparation
Enterobacteria SYA2 is inoculated in the seed culture medium, and under 37 ℃ of temperature, rotating speed 150 r/min shaking tables were cultivated 24 hours, obtained seed liquor;
Seed culture medium is formed: lactose 10 g/L, peptone 10 g/L, yeast extract 3 g/L, sodium-chlor 3 g/L, and pH is 7.0;
B, fermented liquid preparation
Seed liquor by 3% inoculum size, is added in the liquid fermentation medium, and shaking culture is 24 hours under 37 ℃ of temperature, rotating speed 180～200r/min, obtains fermented liquid;
Liquid fermentation medium is formed: lactose 15g/L, peptone 20 g/L, yeast extract 3 g/L, sodium-chlor 3 g/L, dipotassium hydrogen phosphate 1 g/L, Manganse Dioxide 0.1g/L, and pH is 7.0;
Centrifugal and the cytoclasis of C, fermented liquid
With fermented liquid centrifugal 20min under 4 ℃ of temperature, rotating speed 6000r/min condition, collect thalline and be 6.5 phosphate buffer soln washed twice with the pH of 0.1mol/L, behind the thalline ultrasonication 15min, centrifugal 20min under 4 ℃ of temperature, rotating speed 12000r/min obtains supernatant liquor;
The fractionation precipitation of D, enzyme
Ammonium sulfate is dried porphyrize, get the supernatant liquor of C step, reach 35% to wherein slowly adding ammonium sulfate powder to its saturation ratio, after leaving standstill 2.5 hours under 4 ℃ of temperature, with 7000 rev/mins of centrifugal 10min, get supernatant liquor, continue to add ammonium sulfate powder to its saturation ratio and reach 75%, after leaving standstill 2 hours under 4 ℃ of temperature, with 7500 rev/mins of centrifugal 10min, the pH that precipitation is dissolved in 0.01moI/L is in 6.8 the phosphate buffered saline buffer, and dialysis 24 hours under 4 ℃ of temperature again is dialyzate;
The purifying of E, enzyme
Dialyzate is added DEAE-Sephadex A-25 ion exchange column, is that 6.2 phosphate buffered saline buffer is initial damping fluid with the pH of 0.05mo1/L, adopts the sodium-chlor gradient elution, distributes and collects, and detection of active is associated with active elutriant;
The membrane filtration of F, enzyme concentrates
With activated elutriant molecular weight cut-off is that 20,000 daltonian ultra-filtration membranes concentrate 10 times, obtains concentrated lactase liquid;
The lyophilize of G, enzyme
In concentrated lactase liquid, add lyophilized vaccine, the additional proportion of lyophilized vaccine adds the 30g lyophilized vaccine by every 100mL concentrated lactase liquid, with precooling under subzero 20 ℃ of temperature 8 hours, keep temperature of charge to descend dry 24 hours then at subzero 30 ℃, obtain Sumylact L; The weight ratio of lyophilized vaccine consists of, glycine: cyclodextrin: N.F,USP MANNITOL: sodium-chlor=1:1:1:1.
H, pulverizing, packing
To pack after the Sumylact L pulverizing.
Bacterium is compared with mould and yeast, has characteristics such as volume is little, breeding is fast, fermenting process is easy to control, and being applied to suitability for industrialized production has very big potentiality.The bacterium Sumylact L that can be applied to milk-product processing of the present invention's exploitation has certain economic benefits and social benefit.
Morphological specificity and the physiological and biochemical property of enterobacteria SYA2 of the present invention are as follows:
1) morphological specificity: the single bacterium colony of bacterial strain SYA2 is rounded, colourless, projection, smooth surface, translucent.Gramstaining is negative, thalline blunt round, single arrangement, no gemma.Bacterial strain SYA2 bacterium colony and thalli morphology are seen Fig. 1 and Fig. 2 respectively.
2) physiological and biochemical property sees Table 1:
Table 1
| Physiological and biochemical test | The result |
| Glucose | Produce sour aerogenesis |
| Wood sugar | Produce sour aerogenesis |
| N.F,USP MANNITOL | Produce sour aerogenesis |
| Lactose | Produce sour aerogenesis |
| Sodium-chlor | Can grow among 7 % |
| Methyl red | Negative |
| The VP test | Positive |
| The H2S test | Negative |
| Gelatin hydrolysis | Negative |
| Nitrate reduction test | Positive |
3) the enterobacteria SYA2 that makes up according to the 16S rDNA sequence homology of enterobacteria SYA2 sees Fig. 3 with relevant bacterial strain systematic evolution tree.
Enterobacteria of the present invention (
Enterobacter sp) SYA2, in on 09 13rd, 2011 in China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC) preservation, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, preserving number CGMCC No:5251.
Description of drawings
Fig. 1 is the colonial morphology figure of enterobacteria SYA2.
Fig. 2 is the thalli morphology figure of enterobacteria SYA2.
Fig. 3 is enterobacteria SYA2 and the figure of relevant bacterial strain systematic evolution tree.
Embodiment
The following examples can further specify the present invention, but do not limit the present invention in any way.
Embodiment 1
Enterobacteria SYA2 is inoculated in the seed culture medium, and 37 ℃, 150 r/min shaking tables were cultivated 24 hours, obtained seed liquor, and the seed culture medium composition is: lactose 10 g/L, peptone 10 g/L, yeast extract 3 g/L, sodium-chlor 3 g/L, and pH is 7.0;
With seed liquor by 3% inoculum size, add in the liquid fermentation medium, 37 ℃, 180r/min shaking culture 24 hours, obtain fermented liquid, liquid fermentation medium is formed: lactose 15g/L, peptone 20 g/L, yeast extract 3 g/L, sodium-chlor 3 g/L, dipotassium hydrogen phosphate 1 g/L, Manganse Dioxide 0.1g/L, and pH is 7.0;
With fermented liquid centrifugal 20min under 4 ℃, 6000r/min condition, collect thalline and with 0.1mol/L phosphate buffer soln (pH 6.5) washed twice, thalline ultrasonication 15min, the centrifugal 20min of 12000r/min, obtains supernatant liquor by 4 ℃.
Ammonium sulfate is dried porphyrize, get the supernatant liquor that the 3rd step obtained, reach 35% to wherein slowly adding ammonium sulfate powder to its saturation ratio, 4 ℃ leave standstill 2.5 hours after, with 7000 rev/mins of centrifugal 10min, get supernatant liquor, continue to add ammonium sulfate powder to its saturation ratio and reach 75%, 4 ℃ leave standstill 2 hours after, with 7500 rev/mins of centrifugal 10min, precipitation is dissolved in 0.01moI/L, in the phosphate buffered saline buffer of pH6.8, dialysed 24 hours, and be dialyzate for 4 ℃;
Dialyzate is added DEAE-Sephadex A-25 ion exchange column, and with 0.05mo1/L, the phosphate buffered saline buffer of pH 6.2 is initial damping fluid, adopts the sodium-chlor gradient elution, distributes and collects, and detection of active is associated with active eluant,
With activated elutriant, be that 20,000 daltonian ultra-filtration membranes concentrate 10 times with molecular weight cut-off, obtain concentrating enzyme liquid;
In concentrated lactase liquid, add lyophilized vaccine; the additional proportion of lyophilized vaccine is pressed every 100mL concentrated lactase liquid adding 30g lyophilized vaccine; with precooling under-20 ℃ of temperature 8 hours; keep temperature of charge to descend dry 24 hours then at subzero 30 ℃; obtain Sumylact L; enzyme activity reaches 21220U/g, is finished product after packing after Sumylact L is pulverized.
Embodiment 2
Enterobacteria SYA2 is inoculated in the seed culture medium, and 37 ℃, 150 r/min shaking tables were cultivated 24 hours, obtained seed liquor, and the seed culture medium composition is: lactose 10 g/L, peptone 10 g/L, yeast extract 3 g/L, sodium-chlor 3 g/L, and pH is 7.0;
With seed liquor by 3% inoculum size, add in the liquid fermentation medium, 37 ℃, 190r/min shaking culture 24 hours, obtain fermented liquid, liquid fermentation medium is formed: lactose 15g/L, peptone 20 g/L, yeast extract 3 g/L, sodium-chlor 3 g/L, dipotassium hydrogen phosphate 1 g/L, Manganse Dioxide 0.1g/L, and pH is 7.0;
With fermented liquid centrifugal 20min under 4 ℃, 6000r/min condition, collect thalline and with 0.1mol/L phosphate buffer soln (pH 6.5) washed twice, thalline ultrasonication 15min, the centrifugal 20min of 12000r/min, obtains supernatant liquor by 4 ℃.
Ammonium sulfate is dried porphyrize, get the supernatant liquor that the 3rd step obtained, reach 35% to wherein slowly adding ammonium sulfate powder to its saturation ratio, 4 ℃ leave standstill 2.5 hours after, with 7000 rev/mins of centrifugal 10min, get supernatant liquor, continue to add ammonium sulfate powder to its saturation ratio and reach 75%, 4 ℃ leave standstill 2 hours after, with 7500 rev/mins of centrifugal 10min, precipitation is dissolved in 0.01moI/L, in the phosphate buffered saline buffer of pH6.8, dialysed 24 hours, and be dialyzate for 4 ℃.
Dialyzate is added DEAE-Sephadex A-25 ion exchange column, with 0.05mo1/L, the phosphate buffered saline buffer of pH 6.2 is initial damping fluid, adopt the sodium-chlor gradient elution, distribute and collect detection of active, be associated with active eluant, with activated elutriant, be that 20,000 daltonian ultra-filtration membranes concentrate 10 times with molecular weight cut-off, obtain concentrating enzyme liquid; In concentrated lactase liquid, add lyophilized vaccine; the additional proportion of lyophilized vaccine is pressed every 100mL concentrated lactase liquid adding 30g lyophilized vaccine; with precooling under-20 ℃ of temperature 8 hours; keep temperature of charge to descend dry 24 hours then at subzero 30 ℃; obtain Sumylact L; enzyme activity reaches 21280U/g, is finished product after the packing.
Embodiment 3
Enterobacteria SYA2 is inoculated in the seed culture medium, and 37 ℃, 150 r/min shaking tables were cultivated 24 hours, obtained seed liquor, and the seed culture medium composition is: lactose 10 g/L, peptone 10 g/L, yeast extract 3 g/L, sodium-chlor 3 g/L, and pH is 7.0;
With seed liquor by 3% inoculum size, add in the liquid fermentation medium, 37 ℃, 200r/min shaking culture 24 hours, obtain fermented liquid, liquid fermentation medium is formed: lactose 15g/L, peptone 20 g/L, yeast extract 3 g/L, sodium-chlor 3 g/L, dipotassium hydrogen phosphate 1 g/L, Manganse Dioxide 0.1g/L, and pH is 7.0;
With fermented liquid centrifugal 20min under 4 ℃, 6000r/min condition, collect thalline and with 0.1mol/L phosphate buffer soln (pH 6.5) washed twice, thalline ultrasonication 15min, the centrifugal 20min of 12000r/min, obtains supernatant liquor by 4 ℃.
Ammonium sulfate is dried porphyrize, get the supernatant liquor that the 3rd step obtained, reach 35% to wherein slowly adding ammonium sulfate powder to its saturation ratio, 4 ℃ leave standstill 2.5 hours after, with 7000 rev/mins of centrifugal 10min, get supernatant liquor, continue to add ammonium sulfate powder to its saturation ratio and reach 75%, 4 ℃ leave standstill 2 hours after, with 7500 rev/mins of centrifugal 10min, precipitation is dissolved in 0.01moI/L, in the phosphate buffered saline buffer of pH6.8, dialysed 24 hours, and be dialyzate for 4 ℃.
Dialyzate is added DEAE-Sephadex A-25 ion exchange column, with 0.05mo1/L, the phosphate buffered saline buffer of pH 6.2 is initial damping fluid, adopt the sodium-chlor gradient elution, distribute and collect detection of active, be associated with active eluant, with activated elutriant, be that 20,000 daltonian ultra-filtration membranes concentrate 10 times with molecular weight cut-off, obtain concentrating enzyme liquid; In concentrated lactase liquid, add lyophilized vaccine; the additional proportion of lyophilized vaccine is pressed the concentrated enzyme liquid adding of every 100mL 30g lyophilized vaccine; with precooling under-20 ℃ of temperature 8 hours; keep temperature of charge to descend dry 24 hours then at subzero 30 ℃; obtain Sumylact L, enzyme activity is finished product after reaching the 21320U/g packing.
The 16srRNA sequence of enterobacteria SYA2 of the present invention is as follows, altogether 1414bp.
1 TGCAAGTCGA ACGGTAACAG GAAGCAGCTT GCTGCTTTGC TGACGAGTGG CGGACGGGTG
61 AGTAATGTCT GGGAAACTGC CTGATGGAGG GGGATAACTA CTGGAAACGG TAGCTAATAC
121 CGCATAACGT CGCAAGACCA AAGAGGGGGA CCTTCGGGCC TCTTGCCATC GGATGTGCCC
181 AGATGGGATT AGCTAGTAGG TGGGGTAACG GCTCACCTAG GCGACGATCC CTAGCTGGTC
241 TGAGAGGATG ACCAGCCACA CTGGAACTGA GACACGGTCC AGACTCCTAC GGGAGGCAGC
301 AGTGGGGAAT ATTGCACAAT GGGCGCAAGC CTGATGCAGC CATGCCGCGT GTATGAAGAA
361 GGCCTTCGGG TTGTAAAGTA CTTTCAGCGG GGAGGAAGGC GATAAGGTTA ATAACCTTGT
421 CGATTGACGT TACCCGCAGA AGAAGCACCG GCTAACTCCG TGCCAGCAGC CGCGGTAATA
481 CGGAGGGTGC AAGCGTTAAT CGGAATTACT GGGCGTAAAG CGCACGCAGG CGGTCTGTCA
541 AGTCGGATGT GAAATCCCCG GGCTCAACCT GGGAACTGCA TTCGAAACTG GCAGGCTAGA
601 GTCTTGTAGA GGGGGGTAGA ATTCCAGGTG TAGCGGTGAA ATGCGTAGAG ATCTGGAGGA
661 ATACCGGTGG CGAAGGCGGC CCCCTGGACA AAGACTGACG CTCAGGTGCG AAAGCGTGGG
721 GAGCAAACAG GATTAGATAC CCTGGTAGTC CACGCCGTAA ACGATGTCGA CTTGGAGGTT
781 GTGCCCTTGA GGCGTGGCTT CCGGAGCTAA CGCGTTAAGT CGACCGCCTG GGGAGTACGG
841 CCGCAAGGTT AAAACTCAAA TGAATTGACG GGGGCCCGCA CAAGCGGTGG AGCATGTGGT
901 TTAATTCGAT GCAACGCGAA GAACCTTACC TACTCTTGAC ATCCAGAGAA CTTTCCAGAG
961 ATGGATTGGT GCCTTCGGGA ACTCTGAGAC AGGTGCTGCA TGGCTGTCGT CAGCTCGTGT
1021 TGTGAAATGT TGGGTTAAGT CCCGCAACGA GCGCAACCCT TATCCTTTGT TGCCAGCGGT
1081 TAGGCCGGGA ACTCAAAGGA GACTGCCAGT GATAAACTGG AGGAAGGTGG GGATGACGTC
1141 AAGTCATCAT GGCCCTTACG AGTAGGGCTA CACACGTGCT ACAATGGCGC ATACAAAGAG
1201 AAGCGACCTC GCGAGAGCAA GCGGACCTCA TAAAGTGCGT CGTAGTCCGG ATTGGAGTCT
1261 GCAACTCGAC TCCATGAAGT CGGAATCGCT AGTAATCGTG GATCAGAATG CCACGGTGAA
1321 TACGTTCCCG GGCCTTGTAC ACACCGCCCG TCACACCATG GGAGTGGGTT GCAAAAGAAG
1381 TAGGTAGCTT AACCTTCGGG AGGGCGCTTA CCAC
Claims (3)
1. an enterobacteria SYA2 CGMCC No:5251 has the ability of galactopoiesis carbohydrase.
2. the method for preparing Sumylact L with the enterobacteria SYA2 of claim 1 is characterized in that this preparation method is following steps:
A, seed liquor preparation
Enterobacteria SYA2 is inoculated in the seed culture medium, and under 37 ℃ of temperature, rotating speed 150 r/min shaking tables were cultivated 24 hours, obtained seed liquor;
Seed culture medium is formed: lactose 10 g/L, peptone 10 g/L, yeast extract 3 g/L, sodium-chlor 3 g/L, and pH is 7.0;
B, fermented liquid preparation
Seed liquor by 3% inoculum size, is added in the liquid fermentation medium, and shaking culture is 24 hours under 37 ℃ of temperature, rotating speed 180～200r/min, obtains fermented liquid;
Liquid fermentation medium is formed: lactose 15g/L, peptone 20 g/L, yeast extract 3 g/L, sodium-chlor 3 g/L, dipotassium hydrogen phosphate 1 g/L, Manganse Dioxide 0.1g/L, and pH is 7.0;
Centrifugal and the cytoclasis of C, fermented liquid
With fermented liquid centrifugal 20min under 4 ℃ of temperature, rotating speed 6000r/min condition, collect thalline and be 6.5 phosphate buffer soln washed twice with the pH of 0.1mol/L, behind the thalline ultrasonication 15min, centrifugal 20min under 4 ℃ of temperature, rotating speed 12000r/min obtains supernatant liquor;
The fractionation precipitation of D, enzyme
Ammonium sulfate is dried porphyrize, get the supernatant liquor of C step, reach 35% to wherein slowly adding ammonium sulfate powder to its saturation ratio, after leaving standstill 2.5 hours under 4 ℃ of temperature, with 7000 rev/mins of centrifugal 10min, get supernatant liquor, continue to add ammonium sulfate powder to its saturation ratio and reach 75%, after leaving standstill 2 hours under 4 ℃ of temperature, with 7500 rev/mins of centrifugal 10min, the pH that precipitation is dissolved in 0.01moI/L is in 6.8 the phosphate buffered saline buffer, and dialysis 24 hours under 4 ℃ of temperature again is dialyzate;
The purifying of E, enzyme
Dialyzate is added DEAE-Sephadex A-25 ion exchange column, is that 6.2 phosphate buffered saline buffer is initial damping fluid with the pH of 0.05mo1/L, adopts the sodium-chlor gradient elution, distributes and collects, and detection of active is associated with active elutriant;
The membrane filtration of F, enzyme concentrates
With activated elutriant molecular weight cut-off is that 20,000 daltonian ultra-filtration membranes concentrate 10 times, obtains concentrated lactase liquid;
The lyophilize of G, enzyme
In concentrated lactase liquid, add lyophilized vaccine, the additional proportion of lyophilized vaccine adds the 30g lyophilized vaccine by every 100mL concentrated lactase liquid, with precooling under subzero 20 ℃ of temperature 8 hours, keep temperature of charge to descend dry 24 hours then at subzero 30 ℃, obtain Sumylact L;
H, pulverizing, packing
To pack after the Sumylact L pulverizing.
3. the method for preparing Sumylact L as claimed in claim 2 is characterized in that: the weight ratio of lyophilized vaccine consists of among the described step G, glycine: cyclodextrin: N.F,USP MANNITOL: sodium-chlor=1:1:1:1.
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|---|---|---|---|---|
| CN103695507A (en) * | 2013-12-03 | 2014-04-02 | 天津替代医学科技有限公司 | Method for extracting ubiquitin and ubiquitin-like using monoplast mycoprotein |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1837355A (en) * | 2005-07-21 | 2006-09-27 | 山东大学 | Producing microorganism for trans-glycosylation beta-galactosidase |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1837355A (en) * | 2005-07-21 | 2006-09-27 | 山东大学 | Producing microorganism for trans-glycosylation beta-galactosidase |
Non-Patent Citations (3)
| Title |
|---|
| 《Applied Biochemistry and Biotechnology》 20100401 Anamika Ghatak等 beta-D-Galactosidase from Enterobacter cloacae: Production and Some Physicochemical Properties 1678-1688页 1-3 第162卷, 第6期 * |
| 《Biochemical and Biophysical Research Communications》 20070228 Lili Lu等 A novel beta-galactosidase capable of glycosyl transfer from Enterobacter agglomerans B1 78-84页 1-3 第356卷, 第1期 * |
| 《食品科学》 20070531 李正义 等 转糖基beta-半乳糖苷酶生产含低聚半乳糖的低乳糖牛奶 241-244页 1-3 第28卷, 第5期 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103695507A (en) * | 2013-12-03 | 2014-04-02 | 天津替代医学科技有限公司 | Method for extracting ubiquitin and ubiquitin-like using monoplast mycoprotein |
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