NL2030021B1 - L-ARABINOSE ISOMERASE (L-Al) DERIVED FROM LACTOCOCCUS LACTIS (L. LACTIS), AND USE THEREOF IN PREPARATION OF RARE SUGAR - Google Patents

L-ARABINOSE ISOMERASE (L-Al) DERIVED FROM LACTOCOCCUS LACTIS (L. LACTIS), AND USE THEREOF IN PREPARATION OF RARE SUGAR Download PDF

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NL2030021B1
NL2030021B1 NL2030021A NL2030021A NL2030021B1 NL 2030021 B1 NL2030021 B1 NL 2030021B1 NL 2030021 A NL2030021 A NL 2030021A NL 2030021 A NL2030021 A NL 2030021A NL 2030021 B1 NL2030021 B1 NL 2030021B1
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leu
seq
lys
gly
coli
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NL2030021A
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Zhang Susu
Liu Xinli
Wang Ting
Xu Zhenshang
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Univ Qilu Technology
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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)
    • C12N9/2468—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1) acting on beta-galactose-glycoside bonds, e.g. carrageenases (3.2.1.83; 3.2.1.157); beta-agarase (3.2.1.81)
    • C12N9/2471—Beta-galactosidase (3.2.1.23), i.e. exo-(1-->4)-beta-D-galactanase
    • 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
    • C12N9/90—Isomerases (5.)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/24—Preparation of compounds containing saccharide radicals produced by the action of an isomerase, e.g. fructose
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01023—Beta-galactosidase (3.2.1.23), i.e. exo-(1-->4)-beta-D-galactanase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y503/00—Intramolecular oxidoreductases (5.3)
    • C12Y503/01—Intramolecular oxidoreductases (5.3) interconverting aldoses and ketoses (5.3.1)
    • C12Y503/01004—L-Arabinose isomerase (5.3.1.4)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/02—Monosaccharides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/185—Escherichia
    • C12R2001/19—Escherichia coli
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/46—Streptococcus ; Enterococcus; Lactococcus

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Enzymes And Modification Thereof (AREA)

Claims (6)

  1. Conclusies l. Nieuw gen dat codeert voor L-arabinose-isomerase (L-Al), waarbij het nieuwe gen dat codeert voor L-AI een nucleotidesequentie heeft die getoond is in SEQ ID No. 1.
  2. 2. Nieuw L-arabinose-isomerase (L-AD), waarbij het nieuwe L-AI een aminozuursequentie heeft die getoond is in SEQ ID No. 2.
  3. 3. Nieuw gen dat codeert voor L-arabinose-isomerase (L-Al) volgens conclusie 1, waarbij de sequentie aanwezig is in Lactococcus lactis (L. lactis).
  4. 4. Bereidingswerkwijze voor een recombinante stam van Escherichia coli (E. coli), waarbij de werkwijze het volgende omvat: het gebruiken van specifieke primers Fl (5’-GCGCATATGTTAGAAAATACTCAAAAAG-3’) (SEQ ID No. 3) en RI (5-GCGCTCGAGTCATCCAAGATTAATATAAG-3") (SEQ ID No. 4) om het nieuwe gen dat codeert voor L-arabinose-isomerase (L-AT) volgens conclusie 1 te amplificeren; het onderwerpen van PCR-amplificatieproduct aan dubbele enzymdigestie met Neel en Xhol, het lineair maken van een expressievector pETDuet-1 door dezelfde enzymdigestie, en het ligeren van digestieproducten van het amplificatieproduct met de lineair gemaakte expressievector om een recombinante expressievector pETDuet-ara te construeren; het gebruiken van specifieke primers F2 (5-CGCGGATCCGATGAACATGACTGAAAAAATTC-3") (SEQ ID No. 5) en R2 (5’-GCGCTGCAGCTAATTTAGTGGTTCAATCATGAAG-3’) (SEQ ID No. 6) om een B-galactosidase- (B-GAL-) gen van Streptococcus thermophilus (S. thermophilus) te amplificeren; het onderwerpen van een PCR-amplificatieproduct aan dubbele digestie met Bam HI en PST, het lineair maken van de recombinante expressievector pETDuet- ara door dezelfde enzymdigestie, en het ligeren van digestieproducten van het amplificatieproduct met de lineair gemaakte recombinante expressievector om een recombinante expressievector pETDuet-ara-gal te construeren, en het transformeren van de recombinante expressievector pETDuet-ara-gal in een gastheerstam E. coli BL21
    (DE3), het kweken van een transformant, en het induceren van eiwitexpressie middels isopropyl- B-D-thiogalactoside (IPTG) of lactose.
  5. 5. Bereidingswerkwijze van een recombinante stam van Escherichia coli (E. coli) volgens conclusie 4, waarbij de stam lactose direct in D-tagatose om kan zetten.
  6. 6. Bereidingswerkwijze van een recombinante stam van Escherichia coli (E. coli) volgens conclusie 5, waarbij de optimale omstandigheden het volgende omvatten: temperatuur: 50 °C, pH: 8,0 en substraatlactoseconcentratie: 300 g/L.
NL2030021A 2021-12-03 2021-12-03 L-ARABINOSE ISOMERASE (L-Al) DERIVED FROM LACTOCOCCUS LACTIS (L. LACTIS), AND USE THEREOF IN PREPARATION OF RARE SUGAR NL2030021B1 (en)

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NL2030021A NL2030021B1 (en) 2021-12-03 2021-12-03 L-ARABINOSE ISOMERASE (L-Al) DERIVED FROM LACTOCOCCUS LACTIS (L. LACTIS), AND USE THEREOF IN PREPARATION OF RARE SUGAR

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NL2030021B1 true NL2030021B1 (en) 2023-06-21

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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3543334A1 (en) * 2018-03-23 2019-09-25 Danmarks Tekniske Universitet A novel cell-based platform for converting lactose containing feedstocks into value-added compounds

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3543334A1 (en) * 2018-03-23 2019-09-25 Danmarks Tekniske Universitet A novel cell-based platform for converting lactose containing feedstocks into value-added compounds

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
DATABASE EMBL [online] 9 April 2019 (2019-04-09), "Lactococcus lactis strain QL001 L-arabinose isomerase mRNA, complete cds.", XP002807687, retrieved from EBI accession no. EMBL:MH940215 Database accession no. MH940215 *
FARAHAT MOHAMED G: "CODON OPTIMIZATION AND CO-EXPRESSION OF THERMOSTABLE beta-GALACTOSIDASE AND L-ARABINOSE ISOMERASE IN LACTOCOCCUS LACTIS FOR SINGLE-STEP PRODUCTION OF FOOD-GRADE D-TAGATOSE", BIOCHEM. CELL. ARCH., vol. 20, no. 1, 1 April 2020 (2020-04-01), pages 2545 - 2552, XP055968836 *
HERMAN R E ET AL: "CLONING AND EXPRESSION OF THE BETA-D GALACTOSIDASE GENE FROM STREPTOCOCCUS-THERMOPHILUS IN ESCHERICHIA-COLI", APPLIED AND ENVIRONMENTAL MICROBIOLOGY, AMERICAN SOCIETY FOR MICROBIOLOGY, US, vol. 52, no. 1, 1 July 1986 (1986-07-01), pages 45 - 50, XP002283748, ISSN: 0099-2240 *
ZHANG GUOYAN ET AL: "Exploring a Highly D-Galactose Specific L-Arabinose Isomerase From Bifidobacterium adolescentis for D-Tagatose Production", FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY, vol. 8, 29 April 2020 (2020-04-29), XP055968834, DOI: 10.3389/fbioe.2020.00377 *

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