KR100285529B1 - Gene cloning of novel cyclodextrin glucosyltransferase produced from bacillus brevis cd 162 and sequencing of nucleotide sequence - Google Patents

Gene cloning of novel cyclodextrin glucosyltransferase produced from bacillus brevis cd 162 and sequencing of nucleotide sequence Download PDF

Info

Publication number
KR100285529B1
KR100285529B1 KR1019980015898A KR19980015898A KR100285529B1 KR 100285529 B1 KR100285529 B1 KR 100285529B1 KR 1019980015898 A KR1019980015898 A KR 1019980015898A KR 19980015898 A KR19980015898 A KR 19980015898A KR 100285529 B1 KR100285529 B1 KR 100285529B1
Authority
KR
South Korea
Prior art keywords
cyclodextrin
gene
sequencing
nucleotide sequence
novel
Prior art date
Application number
KR1019980015898A
Other languages
Korean (ko)
Other versions
KR19990084279A (en
Inventor
손천배
김명희
오태광
Original Assignee
오태광
김명희
손천배
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 오태광, 김명희, 손천배 filed Critical 오태광
Priority to KR1019980015898A priority Critical patent/KR100285529B1/en
Publication of KR19990084279A publication Critical patent/KR19990084279A/en
Application granted granted Critical
Publication of KR100285529B1 publication Critical patent/KR100285529B1/en

Links

Classifications

    • 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
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/52—Genes encoding for enzymes or proenzymes
    • 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
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/70—Vectors or expression systems specially adapted for E. coli

Landscapes

  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

PURPOSE: Provided are gene cloning of novel cyclodextrin glucosyltransferase produced from bacillus brevis CD 162 and sequencing of nucleotide sequence, thereby producing expensive γ- cyclodextrin in low price, and is thus used in foods, medical supplies, cosmetics, resin products and agricultural medicines. CONSTITUTION: The nucleotide sequence of novel cyclodextrin glucosyltransferase is determined by the following steps of: (a) cloning a gene which codes novel cyclodextrin glucosyltransferase; (b) mapping and subcloning; and (c) sequencing cyclodextrin glucosyltransferase gene.

Description

바실러스 브레비스 CD162(Bacillus brevis CD162)가 생산하는 감마-시클로덱스트린(-cyclodextrin)Gamma-cyclodextrin produced by Bacillus brevis CD162

본 발명은 탐색된 바실러스 브레비스 (Bacillus brevis) 균이 생산하는 감마-시클로덱스트린의 전환율이 높은 신규한 시클로덱스트린 글리코실트랜스퍼라제의 유전자 클로닝 및 유전자 서열 결정에 관한 것이다. 시클로덱스트린는 α-1,4글루코시드 결합에 의해 연결된 6개 이상의 글루코 피라노스 단위로 구성된 환상의 올리고당이다. 12개까지의 글루코스 잔기를 갖는 시클로덱스트린이 알려졌지만, 단지 글루코스가 6개에서 8개로 구성된 세 개의 시클로덱스트린만 현재 산업적으로 생산되고, 그 이용에 대한 연구가 많이 진척되고 있다. 시클로덱스트린는 글루코스 잔기수에 따라 글루코스가 6개이면 알파-시클로덱스트린(α-cyclodextrin), 7개이면 베타-시클로덱스트린(β-cyclodextrin), 8개이면 감마-시클로덱스트린 (γ-cyclodextrin)으로 명명되고 각각의 공동(cavity)의 크기가 174, 262 및 427Å3이다. 또한 시클로덱스트린은 도넛 모양의 독특한 구조를 이루어 수용액에서 시클로덱스트린 분자내 공동의 내부면은 소수성이고 외부면은 친수성이어서 그 공동 내에 여러 유기 및 무기 화합 물질을 포접하는 성질을 갖고 있다(Szejtli, J. Carbonhydrate Polymers. 12:375~392. 1990).이러한 포접작용을 이용 하여 각종 물성을 개선하는 분자 캡슐로 주목되어온 시클로덱스트린는 냄새 가리움(odour masking), 맛 가리움(taste masking) 및 고미 제거, 휘발성 물질의 안정화, 저칼로리 감미료의 감미증진 및 안정화, 광분해성 물질 및 산화물질의 보호, 유화작용, 불안정한 물질의 안정화, 난용성 물질의 가용화, 조해성·접착성 물질의 분말화 등의 효과가 있어 식품, 의약품, 화장품, 수지제품 및 농약 등에 폭넓게 이용되고 있다(Parrish, M. A. Cyclodextrin-a review. 42. 1393. 1989). 특히 감마-시클로덱스트린은 알파-,베타-시클로덱스트린에 비해 공동의 크기가 커서 포접량이 많아 용해도가 큰 특성을 가지고 있다. 그러나 감마-시클로덱스트린은 대량생산하기가 어렵고 단가도 알파-, 베타시클로덱스트린보다 훨씬 높기 때문에 실용화에 많은 제약을 받고 있다. 이러한 문제를 해결하기 위해 많은 연구자들이 감마-시클로덱스트린를 고수율로 생산하는 시클로덱스트린 글리코실트랜스퍼라제 생산 균주의 분리, 유전자의 클로닝과 시클로덱스트린 글리코실트랜스퍼라제의 고발현, 시클로덱스트린 생산을 위한 경제적인 공정의 개발등 시클로덱스트린의 산업적 응용을 만족시킬 수 있는 방법을 모색하기 위해 계속해서 연구하고 있다.The present invention relates to gene cloning and gene sequencing of novel cyclodextrin glycosyltransferase with high conversion of gamma-cyclodextrin produced by the bacterium Bacillus brevis. Cyclodextrins are cyclic oligosaccharides composed of six or more glucopyranose units linked by α-1,4 glucoside bonds. Cyclodextrins with up to 12 glucose residues are known, but only three cyclodextrins consisting of 6 to 8 glucoses are currently produced industrially, and much research is being conducted on their use. Cyclodextrins are named alpha-cyclodextrins by 6 glucose, beta-cyclodextrins by 7, and gamma-cyclodextrins by 8, depending on the number of glucose residues. Each cavity is 174, 262 and 427Å 3 in size. In addition, cyclodextrin has a unique donut-like structure, so that the inner surface of the cyclodextrin intramolecular cavity in the aqueous solution is hydrophobic and the outer surface is hydrophilic so as to enclose various organic and inorganic compounds in the cavity (Szejtli, J. Carbonhydrate Polymers. 12: 375 ~ 392. 1990). Cyclodextrins, which have been noted as molecular capsules for improving various physical properties by using such inclusions, are known for their odor masking, taste masking and bitumen removal, Stabilizes, enhances and stabilizes low-calorie sweeteners, protects photodegradable and oxide materials, emulsifies, stabilizes unstable substances, solubilizes poorly soluble substances, and powders deliquescent and adhesive substances. It is widely used in resins, resin products and pesticides (Parrish, MA Cyclodextrin-a review. 42. 1393. 1989). In particular, gamma-cyclodextrin has a large solubility compared to alpha-, beta-cyclodextrin and has a large solubility so that it has high solubility. However, gamma-cyclodextrin is difficult to mass-produce and the unit cost is much higher than that of alpha- and betacyclodextrin, and thus, it is limited in practical use. In order to solve this problem, many researchers have isolated cyclodextrin glycosyltransferase producing strains producing high yield of gamma-cyclodextrin, cloning of genes and high expression of cyclodextrin glycosyltransferase, and economical methods for producing cyclodextrin. The development of the process continues to explore ways to satisfy the industrial applications of cyclodextrins.

본 발명가들은 시클로덱스트린의 생산의 문제점을 능동적으로 해결하기 위해서 지금까지 보고된 다른 시클로덱스트린 글리코실트랜스퍼라제에 비해 감마-시클로덱스트린을 많이 만드는 신규한 시클로덱스트린 글리코실트랜스퍼라제를 생산하는 균주를 대한민국 특허균주기탁기관인 유전자은행에 기탁(기탁일 : 1998. 4. 22)하여 수탁번호 KCTC 8885P를 부여 받은 바가 있다. 이 특허균주를 이용하여 바실러스 브레비스 CD162의 시클로덱스트린 글리코실트랜스퍼라제의 유전자의 클로닝과 염기 서열 분석을 통해서 유전자를 확보하고, 확보된 유전자를 기준으로 보고된 시클로덱스트린 글리코실트랜스퍼라제와 비교를 통해서 유전자상의 차이점을 인정되어 미국의 NCBI의 GenBank에서 Accession Number (1997년 8월 3일부)를 부여받고 아울러 바실러스 브레비스 CD162의 시클로덱스트린 글리코실트랜스퍼라제의 유전자(pCGMHI164)가 삽입된 대장균을 한국특허균주기탁기관인 유전자 은행에 기탁(기탁일 : 1998년 4월 22일)하여 기탁번호 KCTC 8886P를 부여받았다. 이에 본 발명에서는 신규성이 확인된 시클로덱스트린 글리코실트랜스퍼라제 유전자의 클로닝과 염기 서열 분석을 통해서 유전자를 확보하고, 확보된 유전자를 기준으로 보고된 시클로덱스트린 글리코실트랜스퍼라제와 비교를 통해서 유전자상의 차이점을 확인하여 신규효소임을 확인하고 이효소를 이용한 시클로덱스트린의 생산방법을 확립함에 있다.In order to actively solve the problem of the production of cyclodextrins, the inventors have proposed a Korean patent for a novel cyclodextrin glycosyltransferase that produces more gamma-cyclodextrin compared to other cyclodextrin glycosyltransferases reported so far. It was deposited with the Gene Bank, a strain depositing institution (deposit date: April 22, 1998), and was given accession number KCTC 8885P. Using this patent strain, the gene was obtained by cloning and sequencing the gene of the cyclodextrin glycosyltransferase of Bacillus brevis CD162 and compared with the reported cyclodextrin glycosyltransferase based on the obtained gene. E. coli with the accession number (August 3, 1997) from NCB GenBank of NCBI in the US, and the cyclodextrin glycosyltransferase gene (pCGMHI164) of Bacillus brevis CD162, was inserted. Deposited with the Gene Bank (deposit date: April 22, 1998) was assigned accession number KCTC 8886P. Therefore, in the present invention, the genes are secured through cloning and sequencing of the cyclodextrin glycosyltransferase gene, which has been identified as novel, and compared with the cyclodextrin glycosyltransferase reported on the basis of the secured genes. It confirms that it is a new enzyme and establishes the production method of cyclodextrin using this enzyme.

본 발명은 전분으로부터 감마-시클로덱스트린의 전환률이 9.7%에 달하는 신규 시클로덱스트린 글리코실트랜스퍼라제를 암호화하는 유전자와 그 효소의 염기서열에 관한 것이다. 또한 본 발명은 신규 시클로덱스트린 글리코실트랜스퍼라제의 유전자가 삽입된 E. coli(pCGMH164)가 생산한 효소를 이용하여 전분으로부터 감마-시클로텍스트린을 저렴하게 생산하는데 적용할 수 있다. 이와 같은 본 발명을 더욱 상세히 설명하면 다음과 같다.The present invention relates to a gene encoding a novel cyclodextrin glycosyltransferase with a conversion rate of gamma-cyclodextrin from starch up to 9.7% and a nucleotide sequence of the enzyme. In addition, the present invention can be applied to the low-cost production of gamma-cyclotextrin from starch using an enzyme produced by E. coli (pCGMH164) into which the gene of the novel cyclodextrin glycosyltransferase is inserted. Referring to the present invention in more detail as follows.

[실시예 1] 시클로덱스트린 글리코실트랜스퍼라제 유전자의 클로닝Example 1 Cloning of Cyclodextrin Glycosyltransferase Gene

바실러스 브레비스 CD162 유래의 크로모좀 디엔에이(chromosomal DNA)를 Marmur의 방법(Marmur, J. 1961, J. Mo1. Bio1. 3, 208)에 의해 분리하였다. 분리된 디엔에이를 제한효소인 HindⅢ로 부분절단하고, HindⅢ로 절단한 후 포스파테이즈(phoshpatase, CIP)로 처리한 pUC19과 함께 연결(ligation)시킨 후 감응세포(competent) E. coli JM83에 형질전환(transformation)시켰다. 이것을 엠피실린(ampicillin) 100㎕/㎖ 과 1% 옥수수전분(corn starch)을 첨가한 LB plate에서 37℃에서 16시간 키워 관찰하였다. 약 20,000여주의 형질전환체중 1주가 균체 주위에 투명환을 형성하였다. 투명환을 형성하는 클론(clone)이 시클로덱스트린 글리코실트랜스퍼라제 유전자를 함유하고 있는지를 확인하기 위하여 Kato 등의 방법(Kato, T., and K. Horikoshi. Denpun Kagaku. 1986. 33. 137)에 따라 시클로덱스트린 글리코실트랜스퍼라제 역가 측정을 수행하여 시클로덱스트린 글리코실트랜스퍼라제 유전자가 삽입됐음을 재확인하였다. 이 균을 배양하여 플라스미드(plasmid)를 분리한 결과 3.7 kb의 insert을 함유한 6.4 kb 크기의 플라스미드를 확인하고 pCGMH162로 명명하였다.Chromosomal DNA from Bacillus brevis CD162 was isolated by Marmur's method (Marmur, J. 1961, J. Mo1. Bio1, 3, 208). The isolated DNA was partially cleaved with the restriction enzyme HindIII, digested with HindIII, ligation with pUC19 treated with phosphatase (CIP), and transformed into competent E. coli JM83. (transformation). This was observed for 16 hours at 37 ° C. in an LB plate containing 100 μl / ml ampicillin and 1% corn starch. One week out of about 20,000 transformants formed a clear ring around the cells. In order to check whether the clones forming the transparent ring contain the cyclodextrin glycosyltransferase gene, Kato et al. (Kato, T., and K. Horikoshi. Denpun Kagaku. 1986. 33. 137) Cyclodextrin glycosyltransferase titer measurements were then performed to reconfirm that the cyclodextrin glycosyltransferase gene was inserted. After culturing the plasmid to isolate the plasmid (plasmid) identified a 6.4 kb sized plasmid containing a 3.7 kb insert, named pCGMH162.

[실시예 2] 제한효소지도 (Mapping) 및 SubcloningExample 2 Restriction Enzyme Mapping and Subcloning

6.4 kb의 pCGMH162를 여러 가지 제한효소로 자른 결과, 3.7 kb의 insert DNA안에는 HindⅡ, EcoRⅠ, HindⅢ, PstⅠ, SalⅠ, NdeⅠ, XhoⅠ, XbaⅠ의 제한효소 부위를 가지고 있음을 확인하였다. 효소의 역가를 발현하는데 필요한 유전자만을 찾아내기 위해서 pCGMH164의 subcloning을 행하였다. pCGMH164를 PstⅠ으로 절단하여 self ligation 시켜 E.coli JM83에 형질전환하여 2.7 kb 크기의 insert DNA를 갖는 5.4kb 크기의 pCGMH163을 얻었고, pCGMH163을 다시 SalⅠ과 PstⅠ으로 절단한 후 0.8% 아가로스 젤(agarose gel)에 전기영동하고 Geneclean Ⅱ kit로 5.0 kb DNA를 용출하여 다시 SalⅠ으로 절단하여 self ligation 시켜 E. coli JM83에 형질전환하여 2.3 kb 크기의 insert DHA를 갖는 5.0 kb 크기의 pCGMH164를 얻을 수 있었다(도 1).The 6.4 kb pCGMH162 was cut with various restriction enzymes, and the restriction DNA sites of Hind II, EcoR I, Hind III, Pst I, Sal I, Nde I, Xho I, and Xba I were found in the 3.7 kb insert DNA. Subcloning of pCGMH164 was performed to find only the genes required to express the enzyme titer. pCGMH164 was digested with Pst I and transformed into E. coli JM83 to obtain 5.4 kb sized pCGMH163 with 2.7 kb insert DNA, and pCGMH163 was further digested with Sal I and Pst I and 0.8% agarose gel (agarose) gel) electrophoresed, 5.0 kb DNA was eluted with Geneclean Ⅱ kit, cut again with SalI, self ligation and transformed into E. coli JM83 to obtain 5.0 kb pCGMH164 with 2.3 kb insert DHA. 1).

[실시예 3] 시클로덱스트린 글리코실트랜스퍼라제 유전자의 염기서열 결정Example 3 Determination of the base sequence of a cyclodextrin glycosyltransferase gene

pCGMH164의 2.3 kb 크기의 insert DHA를 sequencing 하기 위해서 제한효소 지도를 근거하여 여러 가지 크기의 deletion subclon을 얻은 후 forward primer와 reverse primer를 이용하여 2.3 kb DHA의 전체 염기 배열을 읽었다. MacMolly 3.5라는 프로그램을 이용하여 2,079 개의 nucleotides(693 amino acid)로 이루어진 시클로덱스트린 글리코실트랜스퍼라제의 open reading frame(ORF)를 찾았으며, Bacillus brevis CD162 균으로부터 분리한 시클로덱스트린 글리코실트랜스퍼라제의 15개의 N-말단 아미노산과 일치함을 확인할 수 있었다(도 2). 본 시클로덱스트린 글리코실트랜스퍼라제는 기존의 Bacillus 속 균주들이 생산하는 시클로덱스트린 글리코실트랜스퍼라제와 다른 염기 서열을 가지는 새로운 시클로덱스트린 글리코실트랜스퍼라제로 생각되며, 아미노산 서열 분석 결과 시클로덱스트린 글리코실트랜스퍼라제의 지질결합자리라고 믿어지는 영역을 포함해 6개의 일치 영역(consensus region)이 있음을 확인하였다.For sequencing the 2.3 kb insert DHA of pCGMH164, we obtained deletion subclon of various sizes based on restriction map and read the entire base sequence of 2.3 kb DHA using forward primer and reverse primer. MacMolly 3.5 was used to find an open reading frame (ORF) of cyclodextrin glycosyltransferase consisting of 2,079 nucleotides (693 amino acids) and 15 cyclodextrin glycosyltransferases isolated from Bacillus brevis CD162. It was confirmed that the N-terminal amino acids match (FIG. 2). The cyclodextrin glycosyltransferase is thought to be a new cyclodextrin glycosyltransferase with a base sequence different from the cyclodextrin glycosyltransferase produced by the strains of the existing Bacillus genus. Six consensus regions were identified, including the region believed to be a lipid binding site.

상술한 바와 같이 본 발명은 고가의 감마-시클로덱스트린을 신규 시클로덱스트린 글리코실트랜스퍼라제 효소를 암호화하는 유전자를 이용해서 효소를 저렴하게 생산할 수 있는 효과가 있다.As described above, the present invention has the effect of producing an expensive gamma-cyclodextrin by using a gene encoding a novel cyclodextrin glycosyltransferase enzyme at low cost.

Claims (4)

신규 시클로덱스트린 글리코실트랜스퍼라제를 암호화하는 아래와 같은 염기서열.The base sequence encoding a novel cyclodextrin glycosyltransferase. 신규 시클로덱스트린 글리코실트랜스퍼라제를 암호화하는 아래 아미노산 서열.The amino acid sequence below which encodes a novel cyclodextrin glycosyltransferase. 청구항 1,2에 의한 염기서열 및 아미노산 서열을 갖는 유전자(pCGMH164)를 함유한 재조합 대장균(수탁번호 KCTC 8886P).Recombinant E. coli (Accession No. KCTC 8886P) containing the gene having the nucleotide sequence according to claims 1 and 2 (pCGMH164). 제3항에 의한 재조합 대장균에 의하여 생산되는 시클로덱스트린 글리코실트랜스퍼라제.Cyclodextrin glycosyltransferase produced by the recombinant Escherichia coli according to claim 3.
KR1019980015898A 1998-05-04 1998-05-04 Gene cloning of novel cyclodextrin glucosyltransferase produced from bacillus brevis cd 162 and sequencing of nucleotide sequence KR100285529B1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
KR1019980015898A KR100285529B1 (en) 1998-05-04 1998-05-04 Gene cloning of novel cyclodextrin glucosyltransferase produced from bacillus brevis cd 162 and sequencing of nucleotide sequence

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
KR1019980015898A KR100285529B1 (en) 1998-05-04 1998-05-04 Gene cloning of novel cyclodextrin glucosyltransferase produced from bacillus brevis cd 162 and sequencing of nucleotide sequence

Publications (2)

Publication Number Publication Date
KR19990084279A KR19990084279A (en) 1999-12-06
KR100285529B1 true KR100285529B1 (en) 2001-05-02

Family

ID=37514407

Family Applications (1)

Application Number Title Priority Date Filing Date
KR1019980015898A KR100285529B1 (en) 1998-05-04 1998-05-04 Gene cloning of novel cyclodextrin glucosyltransferase produced from bacillus brevis cd 162 and sequencing of nucleotide sequence

Country Status (1)

Country Link
KR (1) KR100285529B1 (en)

Also Published As

Publication number Publication date
KR19990084279A (en) 1999-12-06

Similar Documents

Publication Publication Date Title
EP0710674B1 (en) Method for producing a glucan having cyclic structure
Yu et al. The patterned structure of galactoglucomannan suggests it may bind to cellulose in seed mucilage
Kralj et al. Molecular characterization of a novel glucosyltransferase from Lactobacillus reuteri strain 121 synthesizing a unique, highly branched glucan with α-(1→ 4) and α-(1→ 6) glucosidic bonds
Wind et al. Cyclodextrin formation by the thermostable alpha-amylase of Thermoanaerobacterium thermosulfurigenes EM1 and reclassification of the enzyme as a cyclodextrin glycosyltransferase
Gawande et al. Purification and properties of a novel raw starch degrading cyclomaltodextrin glucanotransferase from Bacillus firmus
WO2000014249A1 (en) Nucleic acid molecules encoding an amylosucrase
Meng et al. Biochemical characterization of two GH70 family 4, 6-α-glucanotransferases with distinct product specificity from Lactobacillus aviarius subsp. aviarius DSM 20655
FR2897069A1 (en) CONSTRUCTION OF NEW VARIANTS OF THE DEXTRANE-SACCHARASE DSR-S ENZYME BY MOLECULAR ENGINEERING.
US6248566B1 (en) Glucan having cyclic structure and method for producing the same
WO2010128601A1 (en) Glucuronic acid-containing glucan, process for production of same, and use of same
Rahman et al. Molecular cloning of a cyclodextrin glucanotransferase gene from alkalophilic Bacillus sp. TS1-1 and characterization of the recombinant enzyme
Costa et al. The residue 179 is involved in product specificity of the Bacillus circulans DF 9R cyclodextrin glycosyltransferase
KR100285529B1 (en) Gene cloning of novel cyclodextrin glucosyltransferase produced from bacillus brevis cd 162 and sequencing of nucleotide sequence
Charoensakdi et al. Cloning and expression of cyclodextrin glycosyltransferase gene from Paenibacillus sp. T16 isolated from hot spring soil in northern Thailand
JP3490481B2 (en) Method for producing oligosaccharide
JPH05501059A (en) γ-CGtase
Barreras et al. Functional characterization of GumK, a membrane-associated β-glucuronosyltransferase from Xanthomonas campestris required for xanthan polysaccharide synthesis
JPH10504182A (en) Microorganism capable of synthesizing polyhydroxyalkanoate in a cell simultaneously with synthesis of a polysaccharide extracellularly, and a method for producing the same
Kang et al. Functional, genetic, and bioinformatic characterization of dextransucrase (DSRBCB4) gene in Leuconostoc mesenteroides B-1299CB4
Côté et al. Water-insoluble glucans from sucrose via glucansucrases. Factors influencing structures and yields
Strater et al. Structural basis of the synthesis of large cycloamyloses by amylomaltase
Hyun-Dong et al. Site-directed mutagenesis and functional analysis of maltose-binding site of β-cyclodextrin glucanotransferase from Bacillus firmus var. alkalophilus
US5804426A (en) Recombinant cyclodextran glucanotransferase mutants
Volkova et al. One-step affinity purification of cyclodextrin glucanotransferase from Bacillus sp. 1070
KR19990084278A (en) Novel cyclodextrin glycosyltransferase with high conversion of gamma-cyclodextrin produced by Bacillus brevis CD162 162

Legal Events

Date Code Title Description
A201 Request for examination
E902 Notification of reason for refusal
E701 Decision to grant or registration of patent right
GRNT Written decision to grant
FPAY Annual fee payment

Payment date: 20110201

Year of fee payment: 11

LAPS Lapse due to unpaid annual fee