CN101591628B - Acinetobacter juni. X8 and application thereof in preparing algin lyase - Google Patents

Acinetobacter juni. X8 and application thereof in preparing algin lyase Download PDF

Info

Publication number
CN101591628B
CN101591628B CN2009100995714A CN200910099571A CN101591628B CN 101591628 B CN101591628 B CN 101591628B CN 2009100995714 A CN2009100995714 A CN 2009100995714A CN 200910099571 A CN200910099571 A CN 200910099571A CN 101591628 B CN101591628 B CN 101591628B
Authority
CN
China
Prior art keywords
liquid
acinetobacter
application
sodium alginate
juni
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.)
Expired - Fee Related
Application number
CN2009100995714A
Other languages
Chinese (zh)
Other versions
CN101591628A (en
Inventor
丁玉庭
刘书来
张建友
侯保兵
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Zhejiang University of Technology ZJUT
Original Assignee
Zhejiang University of Technology ZJUT
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Zhejiang University of Technology ZJUT filed Critical Zhejiang University of Technology ZJUT
Priority to CN2009100995714A priority Critical patent/CN101591628B/en
Publication of CN101591628A publication Critical patent/CN101591628A/en
Application granted granted Critical
Publication of CN101591628B publication Critical patent/CN101591628B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Abstract

The invention provides a novel algin lyase producing strain, namely Acinetobacter juni. X8 (which is preserved in the China Center for Type Culture Collection, wherein the preservation number is CCTCCC No: M 209110 and the preservation time is May 20, 2009) and application thereof in preparing algin lyase. The Acinetobacter juni. X8 has the following advantages: (1) the nutritional requirement issimple, the cultivation is easy, the generation time is short, and the fermentation cultivation time is short; (2) intracellular crude enzyme solution is simple to prepare, and a centrifugal collected thallus can be shattered directly after being added into buffer solution; (3) the algin lyase prepared from the Acinetobacter juni. X8 has higher enzyme activity, and unseparated and unpurified crude enzyme solution can reach 53.9 U/mg (157.3 U/mL); (4) the reaction conditions required by the enzyme enzymolysis algin is mild, and the acting time is short; and (5) the algin lyase has good stability, can keep higher enzyme activity (more than 97 percent) after being preserved for 5 days at a temperature of less than or equal to 4 DEG C, and is stable for 8 hours under a condition of which the pH is between 7.7 and 9.0.

Description

Acinetobacter junii X8 and the application in the preparation algin catenase thereof
(1) technical field
The present invention relates to the new algin catenase of a strain and produce bacterium---Acinetobacter junii Acinetobacter juni X8, and the application in the preparation algin catenase.
(2) background technology
Algin is to be present in sea-tangle (Laminaria japonica), sargassun (Gulfweed), linear polysaccharide polymkeric substance in the Sargassum fusiforme frond cell wallss such as (Sargassum fusiforme), be by β-D-1,4-mannuronic acid (β-D-mannuronic acid, be called for short M) and α-L-1,4-guluronic acid (α-L-guluronic acid, abbreviation G) two kinds of acidic polysaccharoses that monomer is formed, along with going deep into of glycobiology and carbohydrate chemistry research, the alginate oligosaccharide of finding different polymerization degree has numerous good biological activity, at new drug, fields such as protective foods exploitation are with a wide range of applications.The alginate oligosaccharide preparation method is multiple, and as acid degradation, alkaline degradation, oxidative degradation, enzymic degradation etc., wherein advantages such as efficient height, specificity are strong, reaction conditions gentleness become from now on main direction of studying because of having for enzyme process preparation.The algin catenase wide material sources, the digestive gland and marine bacteria, the fungi etc. that comprise Sargassum extract, sea mollusk, echinoderms, but because the algin catenase generation is on the low side, separating difficulty is big, storage stability is poor, the application in industrial production still faces a severe challenge.
(3) summary of the invention
The invention provides the new algin catenase of a strain and produce bacterium---Acinetobacter junii (Acinetobacter juni.) X8, and the application in the preparation algin catenase, overcome the shortcoming in the existing algin catenase production preferably.
The technical solution used in the present invention is:
Algin catenase produces bacterium---Acinetobacter junii (Acinetobacter juni.) X8, be preserved in Chinese typical culture collection center, address: China. Wuhan. Wuhan University, 430072, deposit number CCTCCC No:M 209110, preservation date on 05 20th, 2009.
Acinetobacter junii X8 of the present invention obtains by screening in the saprophytic fungus strain of Sargassum fusiforme, screening method is as follows: utilize two kinds of liquid nutrient mediums that bacterial strain is carried out repeatedly enrichment and domestication repeatedly, promptly earlier carry out (30 ℃ of enrichments with 1 pair of bacterial strain screening source of substratum, 180rpm, 72h, the triangular flask of liquid amount 100mL/500mL), screening source microorganism total amount is increased, the back is carried out the bacterial strain selection and is tamed (30 ℃ with 2 pairs of some microorganisms in bacterial screening source of substratum, 180rpm, 72h, the triangular flask of liquid amount 100mL/500mL), carrying out repeatedly so repeatedly, is that the solid medium of sole carbon source carries out separation screening to bacterium producing multi enzyme preparation with sodium alginate again.
The prescription of described two kinds of liquid nutrient mediums is as follows: 1. substratum 1:0.5% sodium alginate, 0.4% peptone, 2.5%NaCl, 1.0%MgSO 47H 2O, 0.2%K 2HPO 43H 2O, 0.001%FeSO 47H 2O, solvent are water, pH value 7.2~7.4; 2. substratum 2:0.5% sodium alginate, 0.5% (NH 4) 2SO 4, 2.5%NaCl, 1.0%MgSO 47H 2O, 0.2%K 2HPO 43H 2O, 0.001%FeSO 47H 2O, solvent are water, pH value 7.2~7.4.
The Acinetobacter junii X8 that the present invention obtains through screening, biological character with stable high yield algin catenase, bacterial strain is after cultivating 48h on the nutrient agar plate, the bacterium colony smooth surface is translucent, and neat in edge is glossy, projection and liquefaction circle are obviously, Gram-positive, shaft-like, morphological specificity as shown in Figure 1, 2.The 16S rDNA sequence of described Acinetobacter junii X8 is as follows:
agagttgatc?atggctcaga?ttgaacgctg?gcggcaggct?taacacatgc?aagtcgagcg
gagatgaggt?gcttgcacct?tatcttagcg?gcggacgggt?gagtaatgct?taggaatctg
cctattagtg?ggggacaaca?ttccgaaagg?aatgctaata?ccgcatacgt?cctacgggag
aaagcagggg?atcttcggac?cttgcgctaa?tagatgagcc?taagtcggat?tagctagttg
gtggggtaaa?ggcctaccaa?ggcgacgatc?tgtagcgggt?ctgagaggat?gatccgccac
actgggactg?agacacggcc?cagactccta?cgggaggcag?cagtggggaa?tattggacaa
tgggggaaac?cctgatccag?ccatgccgcg?tgtgtgaaga?aggccttatg?gttgtaaagc
actttaagcg?aggaggaggc?tactgagact?aatactcttg?gatagtggac?gttactcgca
gaataagcac?cggctaactc?tgtgccagca?gccgcggtaa?tacagagggt?gcgagcgtta
atcggattta?ctgggcgtaa?agcgtgcgta?ggcggctttt?taagtcggat?gtgaaatccc
cgagcttaac?ttgggaattg?cattcgatac?tgggaagcta?gagtatggga?gaggatggta
gaattccagg?tgtagcggtg?aaatgcgtag?agatctggag?gaataccgat?ggcgaaggca
gccatctggc?ctaatactga?cgctgaggta?cgaaagcatg?gggagcaaac?aggattagat
accctggtag?tccatgccgt?aaacgatgtc?tactagccgt?tggggccttt?gaggctttag
tggcgcagct?aacgcgataa?gtagaccgcc?tggggagtac?ggtcgcaaga?ctaaaactca
aatgaattga?cgggggcccg?cacaagcggt?ggagcatgtg?gtttaattcg?atgcaacgcg
aagaacctta?cctggccttg?acatactaga?aactttccag?agatggattg?gtgccttcgg
gaatctagat?acaggtgctg?catggctgtc?gtcagctcgt?gtcgtgagat?gttgggttaa
gtcccgcaac?gagcgcaacc?cttttcctta?cttgccagca?tttcggatgg?gaactttaag
gatactgcca?gtgacaaact?ggaggaaggc?ggggacgacg?tcaagtcatc?atggccctta
cggccagggc?tacacacgtg?ctacaatggt?cggtacaaag?ggttgctaca?cagcgatgtg
atgctaatct?caaaaagccg?atcgtagtcc?ggattggagt?ctgcaactcg?actccatgaa
gtcggaatcg?ctagtaatcg?cggatcagaa?tgccgcggtg?aatacgttcc?cgggccttgt
acacaccgcc?cgtcacacca?tgggagtttg?ttgcaccaga?agtaggtagt?ctaaccgcaa
ggaggacgct?taccacggtg?tggccgatga?ctggggtgaa?gtcgtaacaa?ggtagccgta
ggggaacctg?cggctggatc?acctccttt。
See Fig. 3 according to the strain X 8 of 16S rDNA sequence construct and relevant kind phylogenetic tree thereof.
The invention still further relates to the application of described Acinetobacter junii X8 in the preparation algin catenase.
Concrete, described being applied as: the Acinetobacter junii X8 bacterial classification inoculation after will activating is applicable to the liquid seed culture medium of Acinetobacter junii to the routine that with the sodium alginate is carbon source, obtaining through seed culture that seed liquor is seeded to the sodium alginate is the liquid fermentation medium that the routine of carbon source is applicable to Acinetobacter junii, 28~35 ℃ of fermentation culture 12~28h, acquisition contains the fermented liquid of algin catenase.The gained fermented liquid can carry out separation and purification according to ordinary method, for example through steps such as cytoclasis, ammonium sulfate precipitation, column chromatographies, obtains the algin catenase behind the purifying.Described fermented liquid can be not purified yet, directly applies to the preparation of alginate oligosaccharide, perhaps separates the back is applied to alginate oligosaccharide with somatic cells or immobilized cell form preparation.
Described liquid seeds liquid final concentration is composed as follows: 0.1~0.5% sodium alginate, 0.1~0.5% (NH 4) 2SO 4, 1~5%NaCl, 0.1~0.5%K 4HPO 43H 2O, solvent are water, pH7~8.Among the present invention, substratum is formed all and is represented with quality volume percent (w/v), contains this material 1g in certain concentration of component 1% expression 100mL substratum.
Described liquid fermentation medium final concentration is composed as follows: 0.5~1.0% sodium alginate, 0.5~1.0% peptone, 1~5%NaCl, 0.1~0.5%K 2HPO 43H 2O, solvent are water, pH7~7.5.This nutrient media components is few, and prescription is simple, and yield of enzyme height when thalline utilizes is rare at home and abroad.
Preferably, described application method is as follows:
(1) gets Acinetobacter junii X8 bacterial classification, be seeded to liquid seed culture medium after activated, cultivate 6~10h under 28~32 ℃, 100~200r/min condition, obtain seed liquor; Described liquid seeds liquid final concentration is composed as follows: 0.5% sodium alginate, 0.5% (NH 4) 2SO 4, 2.5%NaCl, 0.2%K 2HPO 43H 2O, solvent are water, pH7.5;
(2) step (1) seed liquor is seeded to liquid fermentation medium with 0.1~10% volume ratio inoculum size, cultivates 16~28h under 25~30 ℃, 100~200r/min condition, obtains containing the fermented liquid of algin catenase; Described liquid fermentation medium final concentration is composed as follows: 0.6% sodium alginate, 0.7% peptone, 1.5%NaCl, 0.3%K 2HPO 43H 2O, solvent are water, pH7.0.
The invention provides and produce algin catenase strains A cinetobacter juni X8, its beneficial effect is mainly reflected in: (1) Acinetobacter junii X8 of the present invention nutritional requirement is simple, cultivate easily, for the time short, fermented incubation time is short; (2) the crude enzyme liquid preparation is simple in the born of the same parents, and centrifugal collection thalline adds directly bacterial cell disruption of damping fluid; (3) this Acinetobacter junii X8 produces the algin catenase enzyme and lives higherly, can reach 53.9U/mg (157.3U/mL) without the crude enzyme liquid of separation and purification; (4) the reaction conditions gentleness of these enzyme enzymolysis algin needs, action time is shorter; (5) a ℃ preservation 5d still keeps high enzyme vigor (>97%) in T≤4 for the algin catenase good stability among the present invention, this algin catenase, and stable in the 8h under the condition of pH7.7~9.0.
(4) description of drawings
Fig. 1 is the colony morphology characteristic of Acinetobacter junii X8 on the isolation medium flat board;
Fig. 2 is the Acinetobacter junii X8 under the oily mirror behind the gramstaining;
Fig. 3 is according to the Acinetobacter junii X8 of 16S rDNA sequence construct and relevant phylogenetic tree of planting thereof.
(5) embodiment
The present invention is described further below in conjunction with specific embodiment, but protection scope of the present invention is not limited in this:
Embodiment 1: the enrichment of bacterial classification, domestication and separation screening
With putrid Sargassum fusiforme leaf and bacterial suspension inoculation to the 500mL triangular flask that fills No. 1 substratum of 100mL, enrichment culture 72h under 30 ℃, 180rpm condition, as well-grown, then pipette the 5mL nutrient solution and be seeded to and fill in No. 2 substratum 30 ℃, 180rpm and cultivate the identical time, cultivate otherwise continue to be seeded in No. 1 substratum.Like this repeated multiple times, moving to then with the sodium alginate is that the substratum of sole carbon source carries out separation screening.
The prescription of described 1, No. 2 liquid and separation screening substratum is as follows:
1. substratum 1:0.5% sodium alginate, 0.4% peptone, 2.5%NaCl, 1.0%MgSO 47H 2O, 0.2%K 2HPO 43H 2O, 0.001%FeSO 47H 2O, solvent are water, pH value 7.2~7.4;
2. substratum 2:0.5% sodium alginate, 0.5% (NH 4) 2SO 4, 2.5%NaCl, 1.0%MgSO 47H 2O, 0.2%K 2HPO 43H 2O, 0.001%FeSO 47H 2O, solvent are water, pH value 7.2~7.4.
3. the separation screening substratum is: 0.5% sodium alginate, 0.5% (NH 4) 2SO 4, 2.5%NaCl, 0.2%K 2HPO 43H 2O, 0.1%MgSO 47H 2O, 0.2% agar, solvent are water, pH value 7.2~7.4.
Through screening the new bacterial strain of the biological character that obtains with stable high yield algin catenase, this bacterial strain is after cultivating 48h on the nutrient agar plate, the bacterium colony smooth surface is translucent, neat in edge, glossy, projection and liquefaction circle are obviously, Gram-positive, shaft-like, morphological specificity such as Fig. 1, shown in 2, through identifying, this bacterial strain is an Acinetobacter junii, called after Acinetobacter junii Acinetobacterjuni.X8 (CCTCCC No:M209110), its 16S rDNA sequence is seen the summary of the invention part, sees Fig. 3 according to the strain X 8 of its 16S rDNA sequence construct and relevant kind phylogenetic tree thereof.
Embodiment 2: the preparation of algin catenase enzyme liquid
The preparation method of algin catenase is as follows:
(1) the strains A cinetobacter juni X8 that will be preserved in slant tube is seeded in the isolation medium flat board, cultivates 48~72h, and the several times of transferring so repeatedly carry out actication of culture; Isolation medium is prepared by following composition: sodium alginate 5g, (NH 4) 2SO 45g, NaCl 25g, K 2HPO 43H 2O2g, MgSO 47H 2O 1g, agar 20g, water complements to 1000mL, pH value 7.2~7.4, heating makes the agar dissolving, cools off dull and stereotypedly;
(2) bacterial classification inoculation after the activation is to the 250mL triangular flask that the 50mL liquid seed culture medium is housed, and 30 ℃, 150rpm cultivates 8h, obtains seed liquor; Described liquid seed culture medium prescription is: 0.5% sodium alginate, 0.5% (NH 4) 2SO 4, 2.5%NaCl, 0.2%K 2HPO 43H 2O, solvent are water, pH7.5;
(3) get the 1.0mL seed liquor and be seeded in the 250mL triangular flask that the 50mL liquid fermentation medium is housed,, cultivate 24h under the 150rpm condition in 25 ℃; The liquid fermentation medium prescription is: 0.6% sodium alginate, 0.7% peptone, 1.5% sodium-chlor, 0.3%K 2HPO 43H 2O, solvent are water, pH7.0.
(4) centrifugal (14500rpm, 4 ℃, 10min) collects thalline;
(5) the distilled water wash bacterial sediment is 3 times, all centrifugal at every turn (14500rpm, 4 ℃, 10min) collecting precipitation;
(6) every 700mL fermented liquid adds the 20mmol/L Tris-HCl damping fluid of 200mL pH 7.5, utilizes ultrasonic disruption instrument smudge cells (power 280W, work/gap 2s/5s, total time 10.5min);
(7) after centrifugal (15400rpm, 4 ℃, 10min) removed cell debris, supernatant liquor was crude enzyme liquid in the born of the same parents (enzyme about 53.9U/mg alive);
(8) 20~60% ammonium sulfate saturation ratios are saltoutd.Concrete grammar is as follows: slowly stir down with magnetic stirring apparatus in the ice bath environment, in born of the same parents, slowly add ammonium sulfate powder to 20% saturation ratio in the crude enzyme liquid and leave standstill 2h, centrifugal (17500rpm, 4 ℃, 10min) getting supernatant liquor continues slowly to add ammonium sulfate to 60% saturation ratio and leave standstill 2h, the centrifugal protein precipitation that gets, after this precipitation is dissolved with 20mmol/LTris-HCl damping fluid (pH 7.5), the desalination of dialysing.
(9) the algin catenase enzyme liquid after saltouing is contained in the dialysis tubing, and the outside concentrates with Macrogol 2000 0 suction.
(10) concentrated solution is filtered with 0.45 μ m millipore filtration (moisture film) after, carry out DEAE-Sepharose F.F chromatography, elutriant is the 20mmol/LTris-HCl damping fluid (pH7.5) of 0.1~0.5mol/L NaCl;
(11) with the 1st active elution peak of DEAE-Sepharose F.F column chromatography, carry out that Macrogol 2000 0 concentrates, after 0.45 μ m millipore filtration (moisture film) filters, last Sephacry S-100 gel column, carry out wash-out with 20mmol/L Tris-HCl damping fluid (pH 7.5), the active elution peak of collection is the enzyme liquid of preparation (enzyme work is about 279.0U/mg).
Embodiment 3: the preparation of alginate oligosaccharide
The alginate oligosaccharide preparation flow is: add the enzyme liquid that 1mL 0.75% (w/w) algin substrate solution, 1mL 0.1moL/L pH7.5 sodium phosphate salt damping fluid, 1mL embodiment 2 make successively in test tube, in 40.0 ℃ of reaction 5min, can be without the alginate oligosaccharide of separation and purification, through 3,5-dinitrosalicylic acid method is measured, and the algin catenase enzymolysis algin behind every mg purifying can about 1395 μ g alginate oligosaccharides.
SEQUENCE?LISTING
<110〉Zhejiang Polytechnical University
<120〉Acinetobacter junii X8 and the application in the preparation algin catenase thereof
<130>
<160>1
<170>PatentIn?version?3.4
<210>1
<211>1529
<212>DNA
<213>Acinetobacter?juni.
<400>1
agagttgatc?atggctcaga?ttgaacgctg?gcggcaggct?taacacatgc?aagtcgagcg 60
gagatgaggt?gcttgcacct?tatcttagcg?gcggacgggt?gagtaatgct?taggaatctg 120
cctattagtg?ggggacaaca?ttccgaaagg?aatgctaata?ccgcatacgt?cctacgggag 180
aaagcagggg?atcttcggac?cttgcgctaa?tagatgagcc?taagtcggat?tagctagttg 240
gtggggtaaa?ggcctaccaa?ggcgacgatc?tgtagcgggt?ctgagaggat?gatccgccac 300
actgggactg?agacacggcc?cagactccta?cgggaggcag?cagtggggaa?tattggacaa 360
tgggggaaac?cctgatccag?ccatgccgcg?tgtgtgaaga?aggccttatg?gttgtaaagc 420
actttaagcg?aggaggaggc?tactgagact?aatactcttg?gatagtggac?gttactcgca 480
gaataagcac?cggctaactc?tgtgccagca?gccgcggtaa?tacagagggt?gcgagcgtta 540
atcggattta?ctgggcgtaa?agcgtgcgta?ggcggctttt?taagtcggat?gtgaaatccc 600
cgagcttaac?ttgggaattg?cattcgatac?tgggaagcta?gagtatggga?gaggatggta 660
gaattccagg?tgtagcggtg?aaatgcgtag?agatctggag?gaataccgat?ggcgaaggca 720
gccatctggc?ctaatactga?cgctgaggta?cgaaagcatg?gggagcaaac?aggattagat 780
accctggtag?tccatgccgt?aaacgatgtc?tactagccgt?tggggccttt?gaggctttag 840
tggcgcagct?aacgcgataa?gtagaccgcc?tggggagtac?ggtcgcaaga?ctaaaactca 900
aatgaattga?cgggggcccg?cacaagcggt?ggagcatgtg?gtttaattcg?atgcaacgcg 960
aagaacctta?cctggccttg?acatactaga?aactttccag?agatggattg?gtgccttcgg 1020
gaatctagat?acaggtgctg?catggctgtc?gtcagctcgt?gtcgtgagat?gttgggttaa 1080
gtcccgcaac?gagcgcaacc?cttttcctta?cttgccagca?tttcggatgg?gaactttaag 1140
gatactgcca?gtgacaaact?ggaggaaggc?ggggacgacg?tcaagtcatc?atggccctta 1200
cggccagggc?tacacacgtg?ctacaatggt?cggtacaaag?ggttgctaca?cagcgatgtg 1260
atgctaatct?caaaaagccg?atcgtagtcc?ggattggagt?ctgcaactcg?actccatgaa 1320
gtcggaatcg?ctagtaatcg?cggatcagaa?tgccgcggtg?aatacgttcc?cgggccttgt 1380
acacaccgcc?cgtcacacca?tgggagtttg?ttgcaccaga?agtaggtagt?ctaaccgcaa 1440
ggaggacgct?taccacggtg?tggccgatga?ctggggtgaa?gtcgtaacaa?ggtagccgta 1500
ggggaacctg?cggctggatc?acctccttt 1529

Claims (6)

1. algin catenase produces bacterium---Acinetobacter junii (Acinetobacter juni.) X8, be preserved in Chinese typical culture collection center, address: China. Wuhan. Wuhan University, 430072, deposit number CCTCC No:M 209110, preservation date on 05 20th, 2009.
2. the application of Acinetobacter junii X8 as claimed in claim 1 in the preparation algin catenase.
3. application as claimed in claim 2, it is characterized in that described being applied as: the Acinetobacter junii X8 bacterial classification inoculation after will activating is the liquid seed culture medium of carbon source with the sodium alginate extremely, being seeded to the sodium alginate through seed culture acquisition seed liquor is the liquid fermentation medium of carbon source, 28~35 ℃ of fermentation culture 12~28h, acquisition contains the fermented liquid of algin catenase.
4. application as claimed in claim 3 is characterized in that described liquid seed culture medium final concentration is composed as follows: 0.1~0.5% sodium alginate, 0.1~0.5% (NH 4) 2SO 4, 1~5%NaCl, 0.1~0.5%K 2HPO 43H 2O, solvent are water, pH7~8.
5. application as claimed in claim 3 is characterized in that described liquid fermentation medium final concentration is composed as follows: 0.5~1.0% sodium alginate, 0.5~1.0% peptone, 1~5%NaCl, 0.1~0.5%K 2HPO 43H 2O, solvent are water, pH7~7.5.
6. application as claimed in claim 2 is characterized in that described application method is as follows:
(1) gets Acinetobacter junii X8 bacterial classification, be seeded to liquid seed culture medium after activated, cultivate 6~10h under 28~32 ℃, 100~200r/min condition, obtain seed liquor; Described liquid seed culture medium final concentration is composed as follows: 0.5% sodium alginate, 0.5% (NH 4) 2SO 4, 2.5%NaCl, 0.2%K 2HPO 43H 2O, solvent are water, pH7.5;
(2) step (1) seed liquor is seeded to liquid fermentation medium with 0.1~10% volume ratio inoculum size, cultivates 16~28h under 25~30 ℃, 100~200r/min condition, obtains containing the fermented liquid of algin catenase; Described liquid fermentation medium final concentration is composed as follows: 0.6% sodium alginate, 0.7% peptone, 1.5%NaCl, 0.3%K 2HPO 43H 2O, solvent are water, pH7.0.
CN2009100995714A 2009-06-11 2009-06-11 Acinetobacter juni. X8 and application thereof in preparing algin lyase Expired - Fee Related CN101591628B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN2009100995714A CN101591628B (en) 2009-06-11 2009-06-11 Acinetobacter juni. X8 and application thereof in preparing algin lyase

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN2009100995714A CN101591628B (en) 2009-06-11 2009-06-11 Acinetobacter juni. X8 and application thereof in preparing algin lyase

Publications (2)

Publication Number Publication Date
CN101591628A CN101591628A (en) 2009-12-02
CN101591628B true CN101591628B (en) 2011-05-25

Family

ID=41406509

Family Applications (1)

Application Number Title Priority Date Filing Date
CN2009100995714A Expired - Fee Related CN101591628B (en) 2009-06-11 2009-06-11 Acinetobacter juni. X8 and application thereof in preparing algin lyase

Country Status (1)

Country Link
CN (1) CN101591628B (en)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102994407B (en) * 2011-12-16 2014-10-29 中国科学院大连化学物理研究所 Flavobacterium strain and incision alginate lyase coding gene, preparation and application
CN104651339B (en) * 2015-02-16 2017-11-03 集美大学 The culture medium and its fermentation process of microvesicle Pseudomonas fermenting and producing algin catenase
CN108530156A (en) * 2018-03-27 2018-09-14 浙江工业大学 A kind of preparation method of alga fertilizer
CN108218587A (en) * 2018-03-27 2018-06-29 浙江工业大学 A kind of compound alga fertilizer of high bioactivity and preparation method thereof
CN112210515B (en) * 2020-10-15 2022-04-26 中国热带农业科学院热带生物技术研究所 Bacterial strain for producing alginate lyase, alginate lyase and application thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1408848A (en) * 2002-09-17 2003-04-09 华东理工大学 Immobile bacillus and method for resolving and preparing chiral cyclopentenylone using said bacillus
CN1920002A (en) * 2005-08-22 2007-02-28 中国海洋大学 Novel vibrionaceae vibrio bacterial strain and application thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1408848A (en) * 2002-09-17 2003-04-09 华东理工大学 Immobile bacillus and method for resolving and preparing chiral cyclopentenylone using said bacillus
CN1920002A (en) * 2005-08-22 2007-02-28 中国海洋大学 Novel vibrionaceae vibrio bacterial strain and application thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
张永 等.琼氏不动杆菌对碳青霉烯类抗生素耐药分子机制研究.《中国抗感染化疗杂志》.2005,第5卷(第2期),77-82. *

Also Published As

Publication number Publication date
CN101591628A (en) 2009-12-02

Similar Documents

Publication Publication Date Title
CN104195080B (en) Bacillus sp capable of producing alginate lyase and application thereof
CN101327975B (en) Method for preparing microorganism flocculant
CN101285046B (en) Mutant strain streptomyces albus TUST2 and process for producing epsilon-polylysine and salts thereof by using the mutant strain
CN101591628B (en) Acinetobacter juni. X8 and application thereof in preparing algin lyase
CN105017086B (en) Separation and purification method for L-citrulline
CN106947724B (en) Method for increasing dissolved oxygen of gamma-polyglutamic acid fermentation liquor
CN102796673A (en) Feruloyl esterase production strain and method for producing feruloyl esterase by using same
CN102154170A (en) Microcystin degrading strain and method for degrading MC-LR (microcystins-LR) by same
CN108018234B (en) Bacterial strain for producing alginate lyase and application thereof
CN102559768A (en) Two-step fermentation production method of microbial flocculant
CN101993847B (en) Bacterial cellulose strain
CN102337299A (en) Preparation method of bacillus flocculant
CN104087628A (en) Method for reducing viscosity of gamma-polyglutamic acid fermentation liquid
CN106278493A (en) The classification enzymatic isolation method preparation method containing oligosaccharide seaweed organic fertilizer
Sun et al. High-efficiency production of Tremella aurantialba polysaccharide through basidiospore fermentation
CN105176859B (en) The bacterial strain MQO-153 of one plant of production arginine deiminase
CN103898013A (en) Thalassospira sp. strain and preparation of kappa-carrageenanase
CN104774794A (en) Strain capable of producing D-mannose isomerase and method for producing D-mannose isomerase by using same
CN101701243B (en) Method for producing R-mandelic acid and derivates thereof by biocatalysis
CN101701202A (en) Enterococcus faecalis and application thereof
CN1253553C (en) Ceramide bacillus and method for production of glycoprotein analog biological flocculant using same
CN1297994A (en) Production process of fungus powder and fungus polysaccharide for food and medicine
CN102965310B (en) Shinella sp. and application thereof to micro-biologically degrading acetaminophen
CN105217799A (en) A kind of industrial fermentation method of molten algae streptomycete active substance
CN105087427B (en) Produce Vibrio natriegen and its application of agarase

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: 20110525

Termination date: 20180611