Ajinomoto Co., Ltd.

Leland Stanford Junior University

Vermillion Corporation

Eiichiro Kimura

Tsuyoshi Nakamatsu

Osamu Kurahashi

Yoshio Kawahara

Kazuhiko Matsui

Shinichi Sugimoto

Yumi Matsuzaki

Yoko Asakura

John P Cooke

Eric Thomas Fung

Jun Nakamura

Seiko Hirano

Gen Nonaka

Tsuyoshi Osumi

Naoki Akiyoshi

Sohei Kanno

Kanae Nakamura

Hisao Ito

Yoshihiro Usuda

Yoshimi Kikuchi

Yasuhiko Yoshihara

Tsuyoshi Ohsumi

Mikiko Suga

Chizu Abe

Shinya Goto

Jun Yamazaki

Nobuharu Tsujimoto

Yosuke Nishio

Akinori Uehara

Sumio Inoue

Takashi Gojobori

Kazuho Ikeo

Yoji Nakamura

Yutaka Kawarabayashi

Yumi Hino

Eiichi Hori

The present invention relates to a method for modifying a property of a protein. The present invention also relates to a method for producing a protein which has a modified property, and a method for producing a microorganism which has a modified property. The present invention is useful in the field of microbial industrial production and the like.

Method for modifying a property of a protein

Disclosed is a biosensor capable of measuring a total concentration of plural types of amino acids. An amino-acid biosensor () for measuring a total concentration of a plurality of specific amino acids, comprises a measuring electrode () which includes as components a mediator and an enzyme which selectively act on the plurality of specific amino acids each serving as a substrate, and a counter electrode (). The enzyme has a substrate affinity to each of the plurality of specific amino acids. The enzyme is operable to catalyze a reaction in each of the plurality of specific amino acids as a substrate so as to form a reaction product, and the mediator is operable, during amino-acid concentration measurement, to carry electrons between the reaction product and the measuring electrode. The amino-acid biosensor is designed to apply a voltage between the measuring electrode and the counter electrode during the measurement in such a manner that, in an analytical curve representing a relationship between an applied voltage and a current value for each of the plurality of specific amino acids, the applied voltage includes a voltage allowing the variety of the current values for the amino acids at the same applied voltage to fall within a given range.

Amino-acid biosensor, fischer-ratio biosensor and health information management system

The present invention provides &#x3b2;2 microglobulin as a biomarker for qualifying or assessing peripheral artery disease in a subject.

Beta-2 microglobulin as a biomarker for peripheral artery disease

Amino-acid biosensor, Fischer-ratio biosensor and health information management system

A method of producing coryneform bacteria having an improved amino acid or nucleic acid-productivity comprises the steps of introducing a mutation in a promoter sequence of amino acid- or nucleic acid-biosynthesizing genes on a chromosome of a coryneform bacterium so that it is close to a consensus sequence or introducing a change in the promoter sequence of amino acid- or nucleic acid-biosynthesizing genes on the chromosome of a coryneform bacterium by gene recombination so that it is close to a consensus sequence, obtaining mutants of the coryneform amino acid- or nucleic acid-producing microorganism, culturing the mutants and select a mutant capable of producing the intended amino acid or nucleic acid in a large amount. This method allows one of skill in the art to construct a mutant capable of enriching or controlling the expression of an intended gene without using a plasmid and to promote production of amino acids in a high yield, by the recombination or mutation.

Method of constructing amino acid producing bacterial strains, and method of preparing amino acids by fermentation with the constructed amino acid producing bacterial strains

A plasmid isolatable from, which comprises a gene coding for a Rep protein having the amino acid sequence shown in SEQ ID NO: 8 or an amino acid sequence having homology of 90% or more to the amino acid sequence shown in SEQ ID NO: 8, and has a size of about 4.4 kb or about 6 kb, or a derivative thereof.

Plasmid autonomously replicable in coryneform bacteria

A plurality of primer sets are designed based on a region where conservation at the amino acid level is observed among various microorganisms for known gene sequences corresponding to a gene coding for an enzyme of the L-amino acid biosynthetic pathway derived from, preferably an enzyme that functions at a higher temperature compared with that of. PCR is performed by using the primers and chromosomal DNA ofas a template. The primers with which an amplification fragment has been obtained are used as primers for screening to select a clone containing a target DNA fragment from a plasmid library of chromosomal DNA of

Genes for heat resistant enzymes of amino acid biosynthetic pathway derived from thermophilic coryneform bacteria

A plurality of primer sets are designed based on a region where conservation at the amino acid level is observed among various microorganisms for known gene sequences corresponding to a gene coding for an enzyme of the L-amino acid biosynthetic pathway derived frompreferably an enzyme that functions at a higher temperature compared with that ofPCR is performed by using the primers and chromosomal DNA ofas a template. The primers with which an amplification fragment has been obtained are used as primers for screening to select a clone containing a target DNA fragment from a plasmid library of chromosomal DNA of

Thermophilic amino acid biosynthesis system enzyme gene of thermotolerant coryneform bacterium

A method of producing coryneform bacteria having improved amino acid or nucleic acid productivity comprising the steps of introducing a mutation in a promoter sequence of amino acid- or nucleic acid-biosynthesizing genes on a chromosome of a coryneform bacterium to make it close to a consensus sequence, or introducing a change in a promoter sequence of amino acid- or nucleic acid-biosynthesizing genes on a chromosome of a coryneform bacterium by gene recombination to make it close to a consensus sequence, to obtain mutants of the coryneform amino acid- or nucleic acid-producing microorganism, culturing the mutants and selecting a mutant capable of producing the intended amino acid or nucleic acid in a large amount. This method allows the construction of a mutant capable of enriching or controlling the expression of an intended gene without using a plasmid and to promote production of amino acids in a high yield by recombination or mutation.

A plasmid which is able to be isolated fromand which comprises a gene coding for a Rep protein having the amino acid sequence shown in SEQ ID NO: 4 or an amino acid sequence having homology of 90% or more to the amino acid sequence shown in SEQ ID NO: 4. and has a size of about 4.4 kb or about 6 kb, or a derivative thereof.

A microorganism is utilized to fermentatively produce useful substances such as amino acids by cultivating the microorganism in a medium to allow a fermentative product to be produced and accumulated in the medium, and collecting the fermentative product, wherein the microorganism to be used is modified by introduction of at least one of a gene coding for a heat shock protein and a gene coding for a &sgr; factor which specifically functions for the heat shock protein gene to enhance expression amount of the heat shock protein in cells, whereby the microorganism is allowed to have added resistance to stress which would otherwise restrain growth of the microorganism and/or production of the fermentative product.

Stress-resistant microorganism and method of producing fermentative product

A coryneform bacterium in which a DNA fragment is incorporated into its chromosome is prepared by (a) obtaining a recombinant plasmid through ligation of a DNA fragment having a sequence homologous to a gene present on a chromosome of a coryneform bacterium to a plasmid that has a wild-type replication control region segment of a particular nucleotide sequence including a mutation and is autonomously replicable in a coryneform bacterium cell at a culture temperature lower than 31&deg; C. but not autonomously replicable in the cell at a temperature of 31&deg; C. or higher, (b) introducing the recombinant plasmid into the coryneform bacterium cell, (c) culturing the bacterium at a temperature of 31&deg; C. or higher, (d) causing homologous recombination between the DNA fragment and the gene present on the chromosome of the coryneform bacterium and having a sequence homologous to the DNA fragment, and (e) selecting a coryneform bacterium in which the DNA fragment is incorporated into its chromosome. According to the present invention, there is provided a method for efficiently modifying genetic traits of a host in a short period of time by obtaining a temperature sensitive plasmid from a plasmid not exhibiting homology with already reported temperature sensitive plasmids or not exhibiting incompatibility therewith.

Temperature sensitive plasmid for coryneform bacteria

Growing community of inventors

Kawasaki, Japan

Eiichiro Kimura