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WO2003018790A1 - Enzyme possedant une activite d'elimination de plaque, sequence d'adn codant pour cette enzyme, cellule hote d'expression et procedes de production et de purification de l'enzyme - Google Patents

Enzyme possedant une activite d'elimination de plaque, sequence d'adn codant pour cette enzyme, cellule hote d'expression et procedes de production et de purification de l'enzyme Download PDF

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Publication number
WO2003018790A1
WO2003018790A1 PCT/KR2001/001439 KR0101439W WO03018790A1 WO 2003018790 A1 WO2003018790 A1 WO 2003018790A1 KR 0101439 W KR0101439 W KR 0101439W WO 03018790 A1 WO03018790 A1 WO 03018790A1
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WO
WIPO (PCT)
Prior art keywords
enzyme
activity
dextranase
amylase
dexamase
Prior art date
Application number
PCT/KR2001/001439
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English (en)
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WO2003018790A8 (fr
Inventor
Doman Kim
Sung Hyuk Kim
Hee Kyoung Kang
Original Assignee
Lifenza Co., Ltd.
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 Lifenza Co., Ltd. filed Critical Lifenza Co., Ltd.
Priority to PCT/KR2001/001439 priority Critical patent/WO2003018790A1/fr
Priority to AU2001286275A priority patent/AU2001286275A1/en
Publication of WO2003018790A1 publication Critical patent/WO2003018790A1/fr
Publication of WO2003018790A8 publication Critical patent/WO2003018790A8/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405—Glucanases
    • C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
    • C12N9/2411—Amylases
    • C12N9/2414—Alpha-amylase (3.2.1.1.)
    • C12N9/2417—Alpha-amylase (3.2.1.1.) from microbiological source
    • C12N9/242—Fungal source
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405—Glucanases
    • C12N9/2451—Glucanases acting on alpha-1,6-glucosidic bonds
    • C12N9/2454—Dextranase (3.2.1.11)

Definitions

  • the present invention relates to an enzyme which can effectively repress formation of plaque or degrade the plaque, gene for encoding the enzyme, and a process for preparing the enzyme and the expressed cell. More specifically, the invention relates to an enzyme which can effectively repress formation of plaque produced from Lipomyces starkeyi KSM 22 mutant and degrade the plaque, gene for encoding the enzyme, a process for producing said enzyme and the expressed cells, and a composition comprising the enzyme.
  • plaque film formed on surface of a tooth (plaque) consists of intimately packed bacteria and non-cellular substances.
  • the main polysaccharide component of plaque is water-insoluble glucan or mutan, which occupies about 20% of total dry weight of plaque, is one of the essential cause of tooth-decay.
  • insoluble glucan essentially comprises ⁇ -1,3-, ⁇ - 1,4- and ⁇ -l,6-D-glucocide bonds.
  • the activity of degrading mutan and that of decomposing starch and dextran are required in order to effectively remove plaque.
  • S. mutans Streptococcus mutans
  • an antibiotic substance to repress growth of S. mutans or a fluorine compound was contained in a product for oral use such as mouth washer.
  • Fluorine as a representative compound of anti-cavity substance, has severe adverse effect of white spot on tooth enamel even at a very low concentration as well as intense toxicity and air pollution, even though it has an effect of repressing growth of bacterial source of tooth-decay.
  • Enzyme such as dextranase has been also used for repressing tooth-decay, but the activity has not been clearly demonstrated yet.
  • U.S. Pat. 5,741,773 provided a toothpaste composition comprising glycomacropeptide having anti-plaque and activity preventing tooth-decay. Such an art, however, simply relates to repression of growth of the bacterial source of tooth-decay, but could not disclose repression of plaque formation or degradation of already formed plaque.
  • the present invention is contrived by said requirements to solve the problems described above.
  • the present invention has an object to provide novel enzyme having activity to repress formation of plaque or to degrade plaque, and gene encoding the enzyme.
  • Another object ofthe present invention is to provide a cell expressing said gene.
  • Still another object of the invention is to provide a process for preparing said enzyme and gene.
  • Still further object of the invention is to provide a commercially useful composition comprised of said enzyme.
  • the present invention provide protein comprising amino acid residue of sequence information 1 or mutants thereof, and enzyme that degrades insoluble glucan having both dextranase activity and amylase activity as a part thereof. Further, the present invention provides said protein or its mutants, gene or mutants of sequence information 2 encoding a part thereof, and/or the segments. Preferably, said mutants of protein comprises not less than 80% of homology with amino acid sequence of sequence information 1, and said gene mutants comprises not less than 80% of homology with gene base sequence of sequence information 2.
  • the present invention also provides transformed cells which express said gene.
  • the cells are preferably prokaryotic cell or eukaryotic cell. The most preferable one is E. coli KCTC 10024BP.
  • the present invention further provides a process preparing a purified enzyme which has both dextranase activity and amylase activity and degrades insoluble glucan, comprising the steps of culturing said cell, expressing an enzyme from the cultured cell, and purifying the expressed enzyme; an enzyme prepared by said process; and a composition comprised ofthe enzyme.
  • Fig. 1 shows the effect of pH on activity and stability of DEXAMase: (a) shows the effect on the activity, while (b) shows the effect on the stability.
  • Fig. 2 shows the effect of temperature on the stability of DEXAMase.
  • Fig. 3 shows a TLC photograph of polysaccharides hydrolyzed by using
  • DEXAMase prepared from recombinant clone pBDEXAM In the photograph, Mn and IMn shows a series of maltodextrin, lane 1 shows dructose, lane 2 shows dextran, lane 3 shows starch, lane 4 shows inulin, lane 5 shows levan, lane 6 shows mutan, lane 7 shows isomaltose, lane 8 shows maltose, lane 9 shows cellular supernatant, lane 10 shows the TLC photograph after reaction with dextran, lane 11 shows one after reaction with starch, lane 12 shows one after reaction with inulin, lane 13 shows one after reaction with levan, lane 14 shows one after reaction with mutan, lane 15 shows one is after reaction with isomaltose, and lane 16 shows one after reaction with maltose.
  • Fig. 4 shows bonding of DEXAMase having the activity of dextranase and amylase on hydroxyapatite.
  • L. starkeyi KSM 22 was cultured in a starch-containing medium, poly(A) + RNA were obtained, cDNA were prepared by the poly(A) + RNA and the cDNA was ligated to Uni-ZAP XR vector.
  • the phage lysate was transformed to pBluescript phagemid via segmenting in vivo after ligation. Then, transformation was subjected thereto by using E. coli XL 1 -Blue to prepare cDNA library.
  • the cDNA library gave clone pBDEXAM formed apparent halo due to decomposition of dextran in a medium containing blue dextran and IPTG. It has been reported that Lipomyces starkeyi (hereinafter, abbreviated to as
  • L. starkeyi produces endodextranase (EC 3.2.1.11) that degrades dextran and ⁇ - amylase that degrades starch.
  • the microorganism is applied in the field of food, but it has not been reported that the microorganism produces antibiotic material or other toxic material.
  • dextranases produced by microorganism are generally inducible enzymes except some bacterial dextranases.
  • the present inventors have disclosed L. starkeyi ATCC 74054 which always produces both dextranase and amylase (U.S. Pat. 5,229,277) with characterizing the properties of the enzyme prepared by the microorganism that produces dextran of small size by using sucrose and starch.
  • the present inventors also has a patent application pending that relates to a microorganism producing dexamase enzyme which can degrade dextran and starch at the same time by using Lipomyces starkeyi KFCC- 11077, an enzyme of the microorganism, and a composition comprised of the enzyme.
  • the enzyme prepared from the gene (dexam) of the present invention degrades not only dextran and starch but also insoluble glucan.
  • the enzyme according to the present invention is referred to as "DEXAM enzyme”.
  • Dexamase of the present invention in case that dextran is used as the substrate, essentially produces glucose, isomaltose and branched tetrose, and small amount of branched pentose and branched hexose.
  • the enzyme When starch is used as the substrate, the enzyme essentially produces glucose, maltose, maltotriose and maltotetrose, and various maltooligosaccharides as well.
  • DEXAM enzyme of the present invention can degrade levan (polymer of ⁇ -fructan), it can effectively degrade iructan, one of the component of plaque formation.
  • DEXAM enzyme of the present invention effectively degrades both soluble glucan and insoluble glucan.
  • the enzyme can repress formation of plaque or remove the plaque already formed by inhibiting aggregation of glucan and bacteria, thereby showing activity on preventing tooth decay.
  • DEXAM enzyme showed the property of retaining the bond to hydroxyapatite, being suggested that DEXAMase can be retained on tooth.
  • the present invention relates to novel microorganism comprising gene encoding DEXAMase.
  • the E. coli strain of the present invention was deposited in KCTC at Yuseong-gu, Daejeon on July 26, 2001 with Deposit No. KCTC 10024BP.
  • the present invention also relates to a process for preparing DEXAM enzyme.
  • the process comprises the steps of culturing pBDEXAM clone and isolating DEXAM enzyme from the culture medium.
  • pBDEXAM can produce DEXAMase by using not only expensive dextran but also glucose, fructose, sucrose or starch, the process is economically advantageous.
  • the properties of DEXAM enzyme obtained from pBDEXAM is substantially identical to those of DEXAMase isolated from L. starkeyi KFCC-11077.
  • the present invention relates to a composition for preventing tooth-decay that comprises DEXAM enzyme.
  • the activity of the enzyme of the present invention can be retained in commercially available mouth washing solutions for a long time, with showing intense tolerance against enzyme inhibitors.
  • the composition containing the enzyme of the present invention can be employed in various oral protective products such as toothpaste and mouth washing solution.
  • the composition comprising the enzyme of the present invention can be applied to food such as chewing gum, soft drink or dairy products.
  • the specific constituents of the composition may be appropriately determined by a person having ordinary skill in the technical field without difficulty. Now, the present invention is described in more detail by referring to Examples which are not limitative:
  • the strain employed as DNA donor for preparing cDNA library was Lipomyces starkeyi KFCC 11077, which constitutively produces DEXAMase having dextranase and amylase activity.
  • XL 1 -Blue and ⁇ -phage Uni-ZAP XR (Stratagene, USA) were employed as host cells and plasmid [phagemid] for preparing cDNA library.
  • L. starkeyi KFCC 11077 was cultured in LMS medium under aerobic condition at 28°C.
  • the LMS medium comprised 1 %(w/v) of starch, l%(v/v) of mineral solution and 0.3%(w/v) of yeast extract.
  • Composition of the mineral solution was 2%(w/v) of MgS0 4 -7H 2 0, 0.1%(w/v) of NaCl, 0.1%( /v) of FeS0 4 -7H 2 0, 0.1%(w/v) of MnS0 4 ⁇ 2 0, and 0.13%(w/v) of CaCl 2 -2H 2 0.
  • LB 1% of tripton, 0.5% of yeast extract, 1% of NaCl, pH 7.3
  • LBA LB containing 50 ⁇ g of ampicillin/ml
  • Example 1 Isolation of poly(A) + RNA from L. starkeyi
  • L. starkeyi KFCC- 11077 was inoculated and cultured at 28°C for 36 hours. At the middle phase of exponential growth, the culture solution was centrifuged (6,500 xg) to obtain a cell pellet. The cell pellet was dissolved in a mixture of GIT solution [4M guanidine isothiocyanate, 25 mM sodium citrate (pH 7.0)], 0.5% lauroylsarcosyl dissolved in distilled water treated with 0.1% DEPC and 0.1 M 2-mercaptoethanol. Glass bead washed with acid and equal amount of phenol (pH 4.0) were added thereto and the mixture was subjected to vortex for 2 minutes. To the supernatant solution obtained after centriftigation, isopropanol was added to precipitate total RNA. From the oligotex-mRNA complex formed, mRNA was isolated by the use of oligotex resin.
  • GIT solution 4M guanidine isothiocyanate, 25 mM sodium citrate (pH
  • cDNA was prepared from poly(A) + RNA (5 ⁇ g) obtained by culturing in a medium containing starch for 36 hours. The prepared cDNA was isolated as the size of not less than 500 bp by spin column separation. Then it was ligated with Uni-ZAP XR vector that had been treated with EcoRI-Xho. The ligated phage cDNA was subjected to packaging in vitro by using Gigapack Gold Kit (Stratagene, U.S.A.). The phage lysate thus obtained was transfered to pBluescript via excision in vivo, transformed with E.
  • the selected colonies are named as pBDEXAM.
  • Plasmid DNA for assay of base sequence was prepared according to alkalinelysis method.
  • the assay of base sequence was performed by means of GeneAmp 9600 thermal cylinder DNA sequencing system (model 373-18, Applied Biosystems, U.S.A.) and ABI PRISM Cycle Sequencing Kit (Perkin Elmer Corp. U.S.A.).
  • the base sequence of DNA fragment (5.4 kb) having glucanhydrolase gene had one open reading frame (ORF) comprised of 2679 bp of bases. Presumed amino acid sequence locates from the starting codon (ATG) at the 874 th nucleotide position ofthe identified base sequence to the terminating codon (TGA) at the 3552 nd position.
  • the structural gene consists of 892 amino acids, and calculated molecular weight is 99.1 kDa.
  • Example 4 Selection of strain which expresses DEXAM gene
  • IPTG isopropyl- ⁇ -D-thiogalactosidase
  • Clone pBDEXAM was cultured in LBA liquid medium at 37°C overnight, of which 1% was inoculated again in LB medium to be cultured to make 0.5 at OD 600nm . IPTG was added until the final concentration became 0.1 mM, and the mixture was cultured at 25 °C overnight. The culture solution was centrifuged (6500 xg, 15 minutes) and the supernatant solution was obtained to be used as the first enzyme solution. In order to concentrate the recovered first enzyme solution, enzyme not less than 30 kDa was concentrated by means of 30K cut-off hallow fiber (Milipore, U.S.A.).
  • DNS 3,5-dinitrosalicylic acid
  • DNS reagent 100 ⁇ l
  • enzyme reaction mixture 50 ⁇ l
  • distilled water 50 ⁇ l
  • absorbance was measured at 550 nm.
  • Dextranase activity of DEXAM enzyme was determined by measuring the amount of isomaltose produced after adding enzyme solution to buffer containing 2% dextran and reacting the mixture at 37°C for 15 minutes.
  • One unit of dextranase is defined by the amount of enzyme that produces 1 ⁇ M of isomaltose after reaction at 37°C for 1 minute by using dextran as substrate.
  • Amylase activity is defined by the amount of enzyme that produces l ⁇ M of maltose after adding the enzyme to buffer containing 2% of starch and reacting the mixture at 37 °C.
  • dextranase activity was 0.16 unit/ml and amylase activity was 0.25 unit/ml.
  • Example 7 Optimal activity and stability of DEXAM enzyme depending on pH or temperature
  • Optimal pH of dextranase and amylase activity of DEXAM enzyme was determined by reaction rate in the pH range of 4 - 9 (by interval of pH 1.0).
  • Optimal temperature of enzyme activity was determined by measuring reaction rate after adding the enzyme at various temperatures (20-90°C, 10°C interval) and standing the mixture for 30 minutes.
  • the temperature stability was obtained by measuring residual activity after placing the enzyme at various temperatures for 30 minutes.
  • Dextranase and amylase of DEXAMase showed optimal activity at pH 5.5.
  • Amylase activity retained not less than 80%o of optimal activity in the range of pH 5.5 - 7, while dextranase activity did in the range of pH 4.0 - 9.0 (Fig. 1, Table 2).
  • the temperature of maximum enzyme activity of dextranase and amylase was 40°C and 50°C, respectively.
  • the dextranase activity and amylase activity retained 80% of initial activity at 50°C or lower temperature and 60°C or lower, respectively. (Fig. 2 and Table 3)
  • Example 8 Degrading properties of DEXAM enzyme on various substrate
  • levan polysaccharide wherein D-fructose is bonded in the position of ⁇ -2,6
  • inulin polysaccharide wherein D-fructose is bonded in the position of ⁇ -2,1
  • mutan polysaccharide wherein D-glucose is bonded in the position of ⁇ - 1,3
  • maltose (4-D- ⁇ -D-glucopyranosyl-D-glucose; disaccharide wherein two moleculues of D- glucose are bonded in the position of ⁇ -1,4) or isomaltose (6-D- ⁇ -D- glucopyranosyl-D-glucose; disaccharide wherein two molecules of D-glucose are bonded in the position of ⁇ - 1,6) was employed.
  • hydroxyapatite which has a lot of identical crystallographic properties to natural apatite existing in bones, is widely used as alternative material in research of artificial bones and teeth.
  • Coherence of DEXAMase to HA was examined. Hydroxyapatite (Bio- Gel HTP, Bio-Rad Laboratories, Richmond CA) was suspended in 10 mM phosphate buffer (pH 6.8). DEXAMase was dissolved in the same buffer solution. The enzyme activity was 10 U/ml. Each 200 ⁇ l of HA and enzyme were mixed, and absorption was performed for 60 minutes.
  • the adsorbed enzyme was gradually eluted by using 10, 50, 100, 200, 300, 400, 500 mM of phosphate buffer (pH 6.8) containing 1 M NaCl. Eluted enzyme fractions are collected, and dextranase and amylase activity was measured.
  • DEXAMase has strong cohesion to hydroxyapatite, so that it remains on teeth for a long time.
  • Ethanol did not affect on activity of each enzyme, and sodium fluoride did not affect on amylase.
  • EDTA reduced about 40% of initial activity of both enzymes, and 0.05% and 0.5% SDS reduced about 35-40% of the activity, while 10-20% ethanol reduced about 45% of dextranase activity and 15-
  • Example 11 Stability of dextranase and amylase against mouth washing components
  • one unit 3 of dextranase or alylase is defined as the amount of enzyme that can produce 1 ⁇ mole of glucolse in an hour by using 2% (w/v) of dextran or water-soluble starch.
  • stable pH range means that residual activity is not less than 60% of the initial activity.
  • stable temperature range means that residual activity is not less than 80% ofthe initial activity.
  • DEXAMase produced by Lipomyces starkeyi KSM 22 according to the present invention is a single protein of about 100 kDa size, and shows both dextranase (EC 3.2.1.11) activity and amylase (EC 3.2.1.1) activity.
  • the base sequence of DNA segment (5.4 kb) had one open reading frame (ORF) comprised of 2679 bp of bases.
  • the gene of presumed structure consists of 892 amino acids, and the molecular weight was about 99.1 kDa.
  • dextranase activity was 0.16 unit/ml, while amylase activity was 0.25 unit/ml.
  • Final reaction products of dextranase and amylase with dextran and starch were typical products of endo-dextranase and endo-amylase.
  • Dextranase activity essentially produced glucose (14.0%), isomaltose (31.2%) and branched tetrasaccharide (33.5%) as well as branched pentasaccharide.
  • Amylase activity essentially produced glucose (28.2%), maltose (18.4%), matotriose (24.7%) and maltotetrase (28.6%).
  • the enzyme was reacted with macromolecular polymers of various structures. As a result, it showed the property of hydrolyzing macromolecular polymer comprising ⁇ -l,3-D-glucoside bond such as mutan as well as iructan comprising ⁇ -bond such as levan and inulin.
  • the microorganism identified under I above was accompanied by:
  • microorganism identified under I above was received by this International Depositary Authority on and a request to convert the original deposit to a deposit under the Budapest Treaty was received by it on

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Abstract

L'invention concerne une enzyme qui inhibe la formation de plaque dentaire et dégrade celle-ci, une séquence d'ADN codant pour l'enzyme, la cellule hôte d'expression, des procédés de production de l'enzyme ainsi que l'enzyme isolée et purifiée. L'invention a également trait à des compositions contenant l'enzyme.
PCT/KR2001/001439 2001-08-25 2001-08-25 Enzyme possedant une activite d'elimination de plaque, sequence d'adn codant pour cette enzyme, cellule hote d'expression et procedes de production et de purification de l'enzyme WO2003018790A1 (fr)

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PCT/KR2001/001439 WO2003018790A1 (fr) 2001-08-25 2001-08-25 Enzyme possedant une activite d'elimination de plaque, sequence d'adn codant pour cette enzyme, cellule hote d'expression et procedes de production et de purification de l'enzyme
AU2001286275A AU2001286275A1 (en) 2001-08-25 2001-08-25 Enzyme with the removal activities of the plaques, dna sequence encoding said enzyme, the expressing host cell and methods for producing and purifying said enzyme

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PCT/KR2001/001439 WO2003018790A1 (fr) 2001-08-25 2001-08-25 Enzyme possedant une activite d'elimination de plaque, sequence d'adn codant pour cette enzyme, cellule hote d'expression et procedes de production et de purification de l'enzyme

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005073369A1 (fr) * 2004-01-30 2005-08-11 Lifenza Co., Ltd. Proteine presentant une activite d'hydrolyse d'amylopectine, d'amidon, glycogene et amylose, gene codant cette proteine, cellule exprimant cette proteine et procede pour la produire
WO2005073368A1 (fr) * 2004-01-30 2005-08-11 Lifenza Co., Ltd. Proteine presentant une activite d'hydrolyse de dextrane, d'amidon, de mutane, d'inuline et de levane, gene codant cette proteine, cellule exprimant cette proteine et procede pour la produire
WO2010018167A1 (fr) * 2008-08-12 2010-02-18 University Of Copenhagen Formulations dentaires destinées à la prévention de l’érosion dentaire
CN105420214A (zh) * 2016-01-26 2016-03-23 华东理工大学 一种α-淀粉酶及其编码基因与应用

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001066570A1 (fr) * 2000-03-09 2001-09-13 Doman Kim Enzyme d'hydrolisation de la plaque dentaire, micro-organisme produisant celle-ci et composition comprenant l'enzyme

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001066570A1 (fr) * 2000-03-09 2001-09-13 Doman Kim Enzyme d'hydrolisation de la plaque dentaire, micro-organisme produisant celle-ci et composition comprenant l'enzyme

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
KELLY C.T. ET AL.: "Thermostable extracellular alpha-amylase and alpha-glucosidase of lipomyces starkeyi", APPL. MICROBIOL. BIOTECHNOL., vol. 22, no. 5, 1985, pages 352 - 358 *
KIM D. ET AL.: "Characterization of a novel carbohydrase from lipomyces starkeyi KSM 22 for dental application", J. MICROBIOL. BIOTECHNOL., vol. 9, no. 3, 1999, pages 260 - 264, XP002468939 *
KOENIG D.W. ET AL.: "Induction of lipomyces starkeyi dextranase", APPL. ENVIRON. MICROBIOL., vol. 55, no. 8, 1989, pages 2079 - 2081 *
RYU S.J. ET AL.: "Purification and partial characterization of a novel glucanhydrolase from lypomyces starkeyi KSM 22 and its use for inhibition of insoluble glucan formation", BIOSCI. BIOTECHNOL. BIOCHEM., vol. 64, no. 2, 2000, pages 223 - 228 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005073369A1 (fr) * 2004-01-30 2005-08-11 Lifenza Co., Ltd. Proteine presentant une activite d'hydrolyse d'amylopectine, d'amidon, glycogene et amylose, gene codant cette proteine, cellule exprimant cette proteine et procede pour la produire
WO2005073368A1 (fr) * 2004-01-30 2005-08-11 Lifenza Co., Ltd. Proteine presentant une activite d'hydrolyse de dextrane, d'amidon, de mutane, d'inuline et de levane, gene codant cette proteine, cellule exprimant cette proteine et procede pour la produire
WO2010018167A1 (fr) * 2008-08-12 2010-02-18 University Of Copenhagen Formulations dentaires destinées à la prévention de l’érosion dentaire
CN105420214A (zh) * 2016-01-26 2016-03-23 华东理工大学 一种α-淀粉酶及其编码基因与应用

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WO2003018790A8 (fr) 2004-04-08

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