SEARCH

Search Details

Kumura Haruto

Field Science Center for Northern BiosphereProfessor

Researcher basic information

■ Degree
  • 博士（農学）, 北海道大学
■ URL
researchmap URLホームページURL■ Various IDs
J-Global ID■ Research Keywords and Fields
Research Keyword
  • 微生物
  • 食品
Research Field
  • Life Science, Food sciences
■ Educational Organization

Research activity information

■ Papers
■ Other Activities and Achievements
■ Books and other publications
  • 乳肉卵の機能と利用（新版）
    玖村 朗人
    アイ・ケイコーポレーション, Sep. 2018
  • 畜産物利用学
    文永堂, 2011
  • 最新畜産物利用学
    朝倉書店, 2006
  • 乳肉卵の機能と利用
    玖村 朗人
    アイ・ケイコーポレーション, 2005
■ Syllabus
  • 大学院共通授業科目（一般科目）：自然科学・応用科学, 2024年, 修士課程, 大学院共通科目
  • 食品科学特論, 2024年, 修士課程, 農学院
  • 食品科学特論演習, 2024年, 修士課程, 農学院
  • 畜産食品衛生学, 2024年, 学士課程, 農学部
  • 基礎畜産物利用学Ⅰ, 2024年, 学士課程, 農学部
  • 畜産科学概論, 2024年, 学士課程, 農学部
  • 応用食品科学, 2024年, 学士課程, 農学部
  • 応用食品科学実験, 2024年, 学士課程, 農学部
■ Affiliated academic society
  • 北海道畜産草地学会
  • 日本畜産学会
  • 日本栄養・食糧学会
  • 日本農芸化学会
■ Research Themes
  • Development of novel food materials for lifestyle disease prevention using edible fungi
    Grants-in-Aid for Scientific Research
    01 Apr. 2013 - 31 Mar. 2017
    KUMURA HARUTO
    Monascus species, known as red-mold has the ability to produce diverse functional secondary metabolites such as lovastatin, monascin and ankaflavin which are responsible for health benefits including risk reduction of arteriosclerosis. It would be applicable for development of functional foods, however, some strains of Monascus sp. produce nephrotoxin, citrinin. Therefore, strain and culture condition should be carefully selected. In addition to the selection of test strains, we focused on the materials for culture substrate, which should be solid with convenience for preparation, if necessary, capability to add nutritional supplements, applicability to wide pH range and “ready to eat” property of the culture products. Following the screening of the suitale strain and culture condition, the resulting culture products were fed to experimental animals whether it could exert predicted biological effects.
    Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, 25350120
  • 食用微生物代謝産物の機能性
    2009
    Competitive research funding
  • Fundamental study on yeasts from dairy origin for the novel probiotic application
    Grants-in-Aid for Scientific Research
    2004 - 2006
    KUMURA Haruto
    In recent years, attention is being paid to the effect of fermented food products on the human health, especially probiotic microorganisms including lactic acid bacteria and Bifidobacterium spp. In this study, immunomodulatory effects and ability of promoting growth and improving viability of Bifidobacterium spp. of some yeast isolated from dairy origin were investigated.
    Despite statistical insignificance, oral administration of some yeasts resulted higher level of immunoglobulin A in the large intestine content of mice. Furthermore, spleen cells prepared from ovalbumin (OVA) immunized mice were incubated in the presence or absence of either OVA, lactic acid bacteria or the 12 yeast strains of 6 species. The results showed that the addition of OVA in the culture of spleen cells stimulated production of immunoglobulin E (IgE); conversely the addition of lactic acid bacteria or yeast cells debris to the culture suppressed IgE production. Although the suppression of IgE production and no induction of IFN-gamma were observed by all yeast strains tested; the level of IL-12 in the culture was found to be strain dependent. In addition, oral administration of a yeast and/or lactic acid bacteria to OVA immunized mice was conducted to monitor the serum IgE level. Reduction of serum IgE level in the experimental group was found to be undetectable as compared with control group. Ability of yeast cell to promote growth and improve viability during refrigeration storage of Bifidobacterium spp. was observed by the addition of yeast extracts to skim culture. Factor(s) associated with growth promotion of Bifidobacterium spp. was found to be thermostable because the effect was being existed even after boiling treatment with skim milk for 30 min.
    Yeast isolated from traditional food is considered as safe and an attractive source to design novel functional foods. Further investigation should be conducted f or beneficial application of yeast.
    Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), HOKKAIDO UNIVERSITY, 16500502
  • Research on Glycosylases which Obtained New Functions by Mutation on Catalytic Residue.
    Grants-in-Aid for Scientific Research
    2002 - 2004
    KIMURA Atsuo; MORI Haruhide; OKUYAMA Masayuki
    Glycosylases are enzymes that hydrolyze the glycosidic linkage. These enzymes also catalyze the transglycosidation, in which the glycosyl residue is transferred to the acceptor substrate. The transglycosidation is an important reaction i)to produce oligosaccharides valuable for foods and ii)to synthesize bio-active sugar-chains. Transglycosidation and hydrolysis proceed in the same time, meaning that the substrate for transglycosidation as well as its product(s) is cleaved by hydrolysis even under conditions of transglycosidation. We have studied the reactions of glycosylase, and have found the phenomena that catalyzed the transglycosidation only. In this study, we analyze the mechanism of valuable phenomena and perform their application. The results are summarized as follows. 1)We have determined the catalytic residue of negatively charged by the method using suicide substrate. Mutant enzyme (synthase), of which catalytic residue was replaced, was produce and purified. The enzyme showed no hydrolytic reaction, only catalyzed the synthesis of oligosaccharide(s) from fluoride-substrate and acceptor. Acceptor of aryl glycoside is a good substrate, meaning that the hydrophobic interaction between aryl-group and subsite +2 is important. 2)Mutant enzyme, which recognized the plane-shaped substrate, a mimic compound of reaction intermediate, was constructed, and its ability of oligosaccharide-synthesis was studied. The low production was observed. We changed the substrate concentration, and succeeded in the improvement of yield. Addition of alcohol to reaction mixture was also effective, but the high concentration of alcohol decreased the production of oligosaccharide. We have found a glycosidase resistant for alcohol. Currently, the conversion of alcohol-stable enzyme to mutant enzyme of same type is trying.
    Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), HOKKAIDO UNIVERSITY, 14360043
  • Structure and functional studies of bovine lactoperoxidase
    Grants-in-Aid for Scientific Research
    1999 - 2000
    SHIMAZAKI Kei-ichi; TANAKA Tetsuya; KUMURA Haruto; NAKAMURA Shingo
    The cDNA encoding bovine lactoperoxidase has been expressed in CHO cells. The recombinant lactoperoxidase was secreted as an enzymatically active single chain molecule presenting two immunoreactive forms of 88 kDa and 82 kDa, differing by their glycosylation. recombinant lactoperoxidase exhibited the characteristic absorbance spectrum with a Soret peak at 413 nm. Biochemical properties of bovine lactoperoxidase isolated from milk and recombinant bovine lactoperoxidase expressed by CHO cells were compared. The natural and recombinant lactoperoxidases showed the same conformational features as determined by CD measurements. The α-helix, β-structure and unordered structure content was found to be 17.8%, 54.2% and 28.0% for the natural lactoperoxidase and 18.6%, 50.1% and 31.3% for the recombinant lactoperoxidase, respectively. The microenvironments of aromatic amino acid residues in both lactoperoxidases seemed to be the same, although the CD spectral band due to the Soret band differed slightly. A difference in the pH-dependent spectral changes of absorbance at 413 nm was observed. From a pepsin hydrolysate of lactoperoxidase, a heme-binding peptide was isolated by reverse-phase HPLC and its amino acid sequence was examined. Engineering of recombinant lactoperoxidase into a myeloperoxidase-like molecule was attempted by substituting Gln-376 by Met, a residue known to achieve covalent binding with the heme in myeloperoxidase. However, the resulting bovine lactoperoxidase mutant failed to acquire the peculiar absorbance spectrum and the chlorinating activity of myeloperoxidase, underlining the complex nature of interactions in the heme vicinity.
    Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B)., HOKKAIDO UNIVERSITY, 11694189
  • ブタ乳腺上皮細胞の培養と樹立株の構築
    科学研究費助成事業
    1997 - 1998
    玖村 朗人
    本研究は平成8年度文部省科学研究費補助金を得て開始されたもので、前年度までに細胞の分離・培養条件をほぼ確立し、細胞の分化機能を観察するためのポリクローナル抗体も調製した。昨年度の時点では分離した乳腺細胞の培養はプラスチックシャーレ上で行なっており、RT-PCRによってカゼインの発現が窺われたもののウエスタンブロットではその発現が検出できなかった。
    今年度はまず乳腺細胞の分化により好適といわれているコラーゲン・ゲルを用いる浮遊培養法に改変し、カゼインの発現を詳細に明らかにしようとした。無血清培地にインシュリン、コルチゾール、プロラクチン等の因子を様々に組み合わせて観察した結果、インシュリンの単独添加でもカゼインの発現が認められることがRT-PCRとウエスタンブロットによって明らかになった。他の動物種由来の乳腺細胞のカゼイン発現に必須であると考えられているプロラクチンを添加してもカゼインの発現は増強されなかった。このことから培養したブタ乳腺上皮細胞はin vitroにおいてなお、カゼインの合成能を有していること、そしてそれはこれまで明かにされてきたプロラクチンとは異なる系が関与していることが示唆された。さらに、本研究は遺伝子導入による分化機能を維持した乳腺細胞の株化を目的としていることから、pSVneoを6種類のリポフェクトアミンによって培養した乳腺細胞に導入し長期間G418存在下で細胞が生存しうるかどうかを検討した。その結果、どのリポフェクトアミンを用いた場合からも長期間安定にベクターが機能する細胞を得るまでには至らなかった。使用するブタの履歴を特定化することや遺伝子を導入する方法を再検討することなどが今後の課題である。
    日本学術振興会, 奨励研究(A), 北海道大学, 09760247
  • ブタ乳腺上皮細胞の分離・樹立
    科学研究費助成事業
    1996 - 1996
    玖村 朗人
    本研究課題であるブタ乳腺上皮細胞の分離・樹立を遂行するためには、1)乳腺組織からの上皮細胞の分離・培養条件の検討、2)in vitroにおける乳蛋白質合成能の確認、3)樹立株の構築のために適用する遺伝子導入法の検討ならびにその条件の最適化が不可欠である。ブタ乳腺上皮細胞の分離はこれまで報告例がなく、さらに遺伝子導入時の転換効率を考慮すれば、まずは他の細胞の混入が極力回避される上皮細胞の分離法の確立が望ましい。そこで、その条件を種々検討した結果、以下のようになった。
    ブタ乳腺組織を細切した後、コラゲナーゼ、デイスパーゼ、ヒアルロニダーゼ、インスリン、DNaseを添加した199倍地を用いて37℃で5時間振とう処理した。一旦細胞を遠心分離によって回収し、DNase、プロナーゼを添加した199倍地を用いて37℃1時間処理した。回収した細胞をDNase、BSA、199培養液を加えて浮遊させ、パーコールを用いた密度勾配遠心分離法によって乳腺細胞の層を採取した。さらに、細胞をナイロンメッシュでろ過し、大きな細胞塊を取り除くと共に、その細胞浮遊液を静置しても沈まず分散した細胞も除去した。このようにして得た乳腺細胞の画分をインスリン、EGF、プロゲステロン、グルココルチコイドを単独あるいは併用で実験を行った結果、マウスやラットとは異なり、ブタ乳腺細胞の増殖にはグルココルチコイドが必要であることが明らかとなった。
    細胞のin vitroにおける乳蛋白質の発現は未確認である。しかし、ブタミルクよりα-、β-カゼイン、β-ラクトグロプリンをカラムクロマトグラフィーによって精製し、マウスに免疫した結果、特に力価の高い抗体として抗β-カゼイン血清を得ることができたため、これを用いて分離した細胞の分化能に関する観察が可能であると考えられる。
    日本学術振興会, 奨励研究(A), 北海道大学, 08760253