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METHOD FOR MASS-PRODUCING BACTERIOCHLOROPHYL b, AND PRODUCING STRAIN

Foreign code F150008543
File No. (S2014-0271-N0)
Posted date Nov 25, 2015
Country WIPO
International application number 2015JP054552
International publication number WO 2015125849
Date of international filing Feb 19, 2015
Date of international publication Aug 27, 2015
Priority data
  • P2014-030085 (Feb 19, 2014) JP
Title METHOD FOR MASS-PRODUCING BACTERIOCHLOROPHYL b, AND PRODUCING STRAIN
Abstract Provided is a bacteriochlorophyl b-producing variant obtained by deleting the bciA gene and bchY and/or bchZ gene inherent to a photosynthetic bacterium which produces bacteriochlorophyl a, and performing a manipulation so as to express the bchY and bchZ genes derived from photosynthetic bacterium that produce bacteriochlorophyl b or bacteriochlorophyl g. Also provided is a method for producing bacteriochlorophyl b, which entails culturing said variant in a culture, and recovering bacteriochlorophyl b from the culture obtained.
Outline of related art and contending technology BACKGROUND ART
Toward the wide use of renewable energy, but needs to advance the introduction of solar power generation, so that cost reduction and mass production of solar cells is important. However, the current solar cell as a starting material for high purity silicon is expensive, its stabilization securing Climate also suffers. As an alternative to next-generation solar cell silicon solar cell, similar to the initial step photosynthesis photo-induced electron transfer of the dye using the dye-sensitized solar cell is attracting attention. So far, artificial porphyrin used as in the case as a sensitizing dye, a natural chlorophyl derivatives using the dye-sensitized solar cell have been reported. However, in order to increase the light energy conversion efficiency, having a wavelength longer than an absorption wavelength in the near-infrared region it is desired to extend.
Is bacteriochlorophyll (BChl), non-oxygen-generating type phototrophic bacteria found from a name given to the tetrapyrrole as, type 7 from there to a g, except BChl f so far been found from natural type 6. Among these, BChl a, and is BChl g BChl b, tetrapyrrole ring B, ring at both D C-C double bond β is a single bond which is reduced to the skeleton has . And purple phototrophic bacteria to BChl b is BChl a, is respectively exist BChl g, - the dye non-covalently bound oligopeptide to form protein complex. The absorption spectrum of chlorophyll (), from the long wavelength side Qy, Qx, By, 4 of the two absorption bands but Bx (By, Bx band are collectively also referred to as Soret band), is BChl a/b/g having a skeleton , chlorine skeleton or plant chlorophyl BChl c/d/e compared with large Qy band in the near infrared region and a long wavelength shift. Among them, is BChl b, due to the presence of ethylidene side chain at position C8, a longer wavelength range than BChl a shifted band Qy. Therefore, in the on position - 17 propionate residues BChl b or long-chain hydrocarbon group has been removed b (BChlide b), from which the central metal thereof desorbed (BPheoide b) b a b (BPhe b) or, if a relatively low cost and in large, the light energy conversion efficiency increases and the useful for the production of an optical device or the like.
In producing bacteria BChl a, 8 - a from (8V-Chlide a), 8 - (or , called DVR) by the action of, an ethyl group and a vinyl group is 8 - reduced to a resulting (Chlide a). Then, from Chlide a, BchX, BchY and BchZ 3 made of a two proteins by the action of the oxidoreductase (COR), C7-8 B ring double bond at position 3 - is a single bond and a resulting (3V-BChlide a) (non-patent document 1), 3 - (BchF) succinate further 3 - and by the action of the C3 - position (BchC) acetylated BChlide a after forming, by the action of BchG, propionate residues on position 17 with long chain hydrocarbon group is added, to produce BChl a. On the other hand, DVR BChl g BChl b and producing bacteria is not present, is BchX/Y/Z (COR), chlorine ring B 8V-Chlide a ring of 1 to a conjugated diene, additional reducing 4, C8 - position of the ring catalyst to catalyze a reaction giving an ethylidene group, and BChl b 8V-Chlide a directly from a precursor of the resulting BChlide g BChl g (Patent Document 1). In producing bacteria BChl b, in the same manner as BChl a producing bacteria, and by the action of BchC BchF, C3 - position of the acetylated BChlide g BChlide b after forming, by the action of the resulting BchG BChl b. Is producing bacteria BchF/BchC BChl g is not present, resulting from the action of BChl g BchG BChlide g.
Many purple phototrophic bacteria belonging to the species to produce BChl a, rare species and produce BChl b, producing bacteria is currently known BChl b, slow grow, cannot grow only under an anaerobic condition, it is possible in the genetic manipulation, to producing large amounts of easily BChl b has not been established yet.
Scope of claims (In Japanese)請求の範囲 [請求項1]
 バクテリオクロロフィルaを産生する光合成細菌において、内在のbciA遺伝子並びにbchY及び/又はbchZ遺伝子を欠損し、かつバクテリオクロロフィルbもしくはバクテリオクロロフィルgを産生する光合成細菌由来のbchY及びbchZ遺伝子を発現するように操作された、バクテリオクロロフィルb産生変異株。

[請求項2]
 バクテリオクロロフィルaを産生する光合成細菌がロドバクター属に属する細菌である、請求項1記載の変異株。

[請求項3]
 ロドバクター属に属する細菌が、ロドバクター・スフェロイデス又はロドバクター・カプスラタスである、請求項2記載の変異株。

[請求項4]
 ロドバクター属に属する細菌が、ロドバクター・スフェロイデスである、請求項2記載の変異株。

[請求項5]
 ブラストクロリス・ビリディス由来のbchY及びbchZ遺伝子を発現するように操作された、請求項1~4のいずれか1項に記載の変異株。

[請求項6]
 バクテリオクロロフィルaを産生する光合成細菌に内在するbciA遺伝子並びにbchY及び/又はbchZ遺伝子を欠損させ、さらにバクテリオクロロフィルbもしくはバクテリオクロロフィルgを産生する光合成細菌由来のbchY及びbchZ遺伝子を発現するように、該細菌を操作することを特徴とする、バクテリオクロロフィルb産生変異株の製造方法。

[請求項7]
 請求項1~5のいずれか1項に記載の変異株を培地中で培養し、得られる培養物からバクテリオクロロフィルbを回収することを含む、バクテリオクロロフィルbの製造方法。

  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • THE RITSUMEIKAN TRUST
  • KURUME UNIVERSITY
  • Inventor
  • TSUKATANI, Yusuke
  • TAMIAKI, Hitoshi
  • HARADA, Jiro
  • FUJITA, Yuichi
  • NOMATA, Jiro
IPC(International Patent Classification)
Specified countries National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JP KE KG KN KP KR KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG
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