Micro Biopharm Japan Co., Ltd.

B Food Science Co., Ltd.

National Institute of Advanced Industrial Science and Technology

Tadashi Fujii

Keisuke Tamura

Yoshikazu Fujii

Takumi Tochio

Saki Nakamura

Atsushi Ochiai

Masashi Ito

Kazuhiro Machida

Naomi Ito

Tomohiro Tamura

Yoshimi Fukatani

Misa Yahara

Akira Arisawa

Hiroki Kabumoto

To provide an improved &#x3b2;-fructofuranosidase which is capable of efficiently producing kestose while inhibiting the production of nystose. This improved &#x3b2;-fructofuranosidase comprises either: an amino acid sequence (a) obtained by introducing, into the amino acid sequence represented by SEQ ID NO: 2, an amino acid mutation i) in which the 85th glycine (G) from the N-terminal is substituted for a protein-constituting amino acid other than glycine (G), and/or an amino acid mutation ii) in which the 310th histidine (H) from the N-terminal is substituted for lysine (K), arginine (R), or tyrosine (Y); or an amino acid sequence (b) which exhibits &#x3b2;-fructofuranosidase activity, and which comprises an amino acid sequence obtained by deleting, substituting, inserting, or adding one or more amino acids in (a) other than the amino acid into which the amino acid mutation has been introduced.

&#x3b2;-fructofuranosidase

An improved &#x3b2;-fructofuranosidase is provided. The improved &#x3b2;-fructofuranosidase may comprise an amino acid sequence having 60% or higher identity to the amino acid sequence of SEQ ID NO: 2, and may contain an amino acid mutation that replaces histidine (H) corresponding to position 395 counted from the N terminus of SEQ ID NO: 2 with arginine (R) or lysine (K), an amino acid mutation that replaces leucine (L) at position 123 counted from the N terminus of SEQ ID NO: 2 with cysteine (C), and/or an amino acid mutation that replaces phenylalanine (F) at position 473 counted from the N terminus of SEQ ID NO: 2 with tyrosine (Y).

[Problem] To provide a production method capable of simply and efficiently producing a fructose-added carbohydrate using &#x3b2;-fructofuranosidase. [Solution] A method for producing a fructose-added carbohydrate, said method having a step in which a receptor substrate and a hydrate containing terminal fructose residue are brought into contact with:

Method for producing fructose-added carbohydrate

A method for producing cis-5-hydroxy-L-pipecolic acid is described. A gene recombinant microorganism enabling direct production of cis-5-hydroxy-L-pipecolic acid can be used in the method. Also described is a gene recombinant microorganism. In particular, it is described that a gene recombinant microorganism having DNAs encoding proteins involved in the biosynthesis of L-pipecolic acid and a DNA encoding a protein having the L-pipecolic acid cis-5-hydroxylase activity is cultured in a medium, and cis-5-hydroxy-L-pipecolic acid is collected from the medium.

Biological method for producing cis-5-hydroxy-L-pipecolic acid

An expression vector capable of expressing a foreign gene in; a transformant ofproduced by using the expression vector; a method for producing a protein by using the transformant; a method for producing an active form of vitamin D3 from vitamin D3, which comprises highly expressing a gene encoding an enzyme involved in the synthesis of the active form of vitamin D3 in a transformant by using the expression vector or the transformant; a method for producing 25-hydroxyvitamin D2 from vitamin D2; and a method for producing pravastatin from compactin, which comprises highly expressing a compactin hydroxylase gene in a transformant by using the expression vector or the transformant.

Expression vector for pseudonocardia autotrophica

Disclosed is a means for improving the poor conversion efficiency in a conventional bioconversion system using a transformant which is given by introducing a gene originated from xerogenic organisms. A transformant is prepared by using a host which is defective in a gene encoding a multidrug efflux protein and introducing a gene originated from xerogenic organisms. Use of the transformant results in much effective microbial conversion of a hydrophobic or amphipathic substrate compound into a desired compound. In case, anis used as the host, the gene encoding a multidrug efflux protein to be defective may be tolC, acrA, acrB and the like.

Transformed strains originated from multidrug efflux protein defective strains and a method for microbial conversion using them

Disclosed is a means for improving the poor conversion efficiency in a bioconversion system using ancell having a bacterium-originated cytochrome P-450 gene integrated therein. A recombinantcell is produced by introducing aciB and aciC which encode a gene for the electron transport system originated from thesp. OC4 strain into ancell, and adding a polynucleotide encoding an N-terminal sequence composed of 48 amino acid residues of AciA and the like to the 5'-terminus of a bacterium-originated cytochrome P-450 gene, wherein AciA is an alkane-oxidative cytochrome P-450 originated from thesp. OC4 strain. Use of the recombinantcell results in much effective microbial conversion of a hydrophobic or amphipathic substrate compound into a desired compound.

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Chuo-ku, Japan

Tadashi Fujii