Biosynthesis of Polyhydroxyalkanoates and 5-Aminolevulinic Acid by Rhodopseudomonas sp. KCTC1437

Rhodopseudomonas sp. KCTC1437에서의 Polyhydroxyalkanoates와 5-Aminolevulinic Acid의 생합성

  • Published : 2002.06.01

Abstract

For elucidating the relationship between the biosynthetic pathways for polyhydroxyslkanoates (PHAs) and 5-aminolevulinic acid (ALA), culture conditions for the production of these two biomaterials by Rhodopseudomonas sp. KCTC 1437 were investigated. Of the carbon substrates tested, acetic acid was the best carbon source for cell growth and PHA biosynthesis. When succinic acid was added as a co-substrate into culture medium, cell growth and PHA production were greatly increased up to 2.5 g/ι and 73% of dry cell weight, respectively. The PHA obtained from the carbon substrates tested was homopolyester of 3-hydroxybutyrate, while valeric acid was only effective for the production of copolyester consisting of 3-hydroxybutyrate and 3-hydroxyvalerate. Anaerobic light culture condition was better for PHA production and cell growth than anaerobic dark or aerobic dark culture condition. The organism was capable of synthesizing ALA when glycine and succinic acid were added to the culture medium. ALA was produced to ca.400 mg/ι when levulinic acid, soccinic acid, and glycine were repeatedly added with a reductant (sodim thioglycolate). However, the presence of glycine, levulinic acid and sodium glycolate inhibited the cell growth and the conversion of carbon substrates to PHA. From these results it is apparent that the production yields of PHA and ALA could not be increased simultaneously because the optimal conditions for the production of PHA and ALA are opposed to each other.

Rhodopseudomonas sp. KCTC1437균주를 이용하여 Polyhydroxyalkanoate (PHA)와 5-aminolevulinic acid (ALA)를 생산하기 위한 배양조건과 이들 생합성 조건의 상호관련성에 대하여 조사하였다. PHA생합성을 위한 탄소원으로는 acetic acid가 가장 효과적이었으나, succinic acid를 보조 탄소원으로 사용하였을 때의 세포건체량은 2.5g/ι, PHA함량은 건체량의 73%로서 주 탄소원만을 사용할 때에 비하여 크게 증가하였다. 조사된 탄소원으로부터 생합성된 PHA는 모두 polyhydroxybutyrate 단일중합체 이었으나, valeric acid로부터는 3-hydroxybutyrate와 3-hydroxyvalerate로 구성된 공중합체가 생산되었다. ALA의 생합성을 위하여서는 환원제 인 sodium thioglycolate를 첨가하여 혐기적 조건을 만들고, 탄소원인 acetic acid와 propionic acid 이외에 전구물질인 levulinic acid, succinic acid와 glycine을 반복적으로 공급해주었을 때 가장 좋았으며, 약 400 mg/ι의 ALA를 생산할 수 있었다. 그러나 ALA 생합성의 필수물질인 glycine, levulinic acid와 환원제는 세포생장과 PHA의 생합성을 저해하는 것으로 나타났다. 이러한 실험 결과, Rhodopseudomonas sp. KCTC1437균주로부터 PHA와 ALA를 동시에 생산할 수 있으나, 두 가지 유용산물을 효율적으로 생합성하기 위한 각각의 배양조건이 상호 배타적임을 확인하였다.

Keywords

References

  1. FEMS Microbiol. Rev. v.103 Studies on copolyester synthesis by Rhodococcus ruber and factors influncing the molecular mass of polyhydroxybutyrate accumulated by Methylobacterium extroquens and Alcaligenes eutophus Anderson, A.J.;D.R. Williams;B. Taidi;E.A. Dawes;D.F. Ewing https://doi.org/10.1111/j.1574-6968.1992.tb05826.x
  2. Int. J. Biol. Macromol. v.11 Ability of the phototrophic bacterium Rhodospirllum rubrum to produce various poly(3-hydroxyalkanoates);Potential sources for biodegradable polyesters Brandl, H.;E.J. Knee Jr;R.C. Fuller https://doi.org/10.1016/0141-8130(89)90040-8
  3. J. Biotechnol. v.65 Polyhydroxy-alkanoates, biopolyesters from renewable resources: Physiological and engineering aspects Braunegg, G.;G. Lefebvre;K.F. Genser https://doi.org/10.1016/S0168-1656(98)00126-6
  4. J. Microbiol. v.39 Effect of levulinic acid on the production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) by Ralstonia eutropha KHB-8862 Chung, S.H.;G.G. Choi;H.W. Kim;Y.H. Rhee
  5. J. Ferment. Bioeng. v.86 Effect of oligosaccharides on glucose consumption by Rhodobacter sphaeroides in polyhydroxyal-kanoate production from enzymatically treated crude sago starch Hassan, M.A.;Y. Shirai;A. Kubota;M.I.A. Karim;K. Nakanishi;K. Hashimoto https://doi.org/10.1016/S0922-338X(98)80034-2
  6. Appl. Environ. Microbiol. v.56 Accumulation of a polyhydroxyalkanoate containing primarily 3-hydroxydecanoate from simple carbohydrate substrates by Pseudomonas sp. strain NCIMB 40135 Haywood, G.W.;A.J. Anderson;D.F. Ewing;E.A. Dawes
  7. Biosci. Biotech. Biochem. v.61 New physiological effects of 5-aminolevulinic acid in plants: The increase of photosynthesis chlorophyll content and plant growth Hotta, Y.;T. Yanaka;H. Takoka;Y. Takeuchi;M. Konnai https://doi.org/10.1271/bbb.61.2025
  8. Int. J. Biol. Macromol. v.28 Evaluation of various carbon substrates for the biosynthesis of polyhydroxyalkanoates bearing functional groups by Psudomonas putida Kim, D.Y.;Y.B. Kim;Y.H. Rhee https://doi.org/10.1016/S0141-8130(00)00150-1
  9. Photochem. Photobiol. v.64 Prodrugs of 5-aminolevulinic acid for photodynamic therapy Kloek, J.;G.M.J.B. van Henegouwen https://doi.org/10.1111/j.1751-1097.1996.tb01868.x
  10. J. Microbiol. v.33 Biosynthesis of poly-β-hydroxyalkanoates by Bacillus thuringiensis R-510 Lee, K.T.;J.Y. Kim;Y.H. Rhee;K.S. Bae;Y.B. Kim
  11. Arch. Microbiol. v.155 Formation of poly(3-hydroxyalkanoates) by phototrophic and chemolithotrophic bacteria Liebergesell, M.;E. Hustede;A. Timm;A. Steinbuchel;R.C. Fuller;R.W. Lenz;H.G. Schlegel https://doi.org/10.1007/BF00244955
  12. Microbiol. Mol. Biol. Rev. v.63 Metabolic engineering of poly(3-hydroxyalkanoates): from DNA to plastic Madison, L.L.;G.W. Huisman
  13. J. Biol. Chem. v.219 The occurrence and determination of 5-aminolevulinic acid and porphobilinogen in urine Mauzerall, D.;S. Granick
  14. FEMS Microbiol. Rev. v.103 Factors affecting poly-hydroxybutyrate accumulation in cyanobacteria and in purple non-sulfur bacteria Philippis, R.D.;A. Elba;M. Guastini;C. Sili;M. Vincenzini
  15. Enzyme Microb. Technol. v.6 Photodynamic herbicides: 1. Concept and phenomenology Rebeiz, C.A.;A. Montazar-Zouhoor;H.J. Hopen;S.M. Wu https://doi.org/10.1016/0141-0229(84)90012-7
  16. Biothechnol. Lett. v.19 Inhibition of 5-aminolevulinic acid (ALA) dehydratase by undissociated levulinic acid during ALA extracellular formation by Rhodobacter sphaeroides Sasaki, K.;M. Wasanori;N. Nishio https://doi.org/10.1023/A:1018331824331
  17. J. Ferment. Technol. v.65 Production of 5-aminolevulinic acid by photosynthetic bacteria Sasaki, K.;S. Ikeda;Y. Nishizawa;M. Hayashi https://doi.org/10.1016/0385-6380(87)90109-9
  18. Appl. Microbiol. Biotechnol. v.32 Production of a herbicide, 5-aminolevulinic acid, by Rhodobacter sphaeroides using the effluent of swine waste from an anaerobic digestor Sasaki, K.;T. Tanaka;Y. Nishizawa;M. Hayashi https://doi.org/10.1007/BF00164749
  19. Biotechnol. Prog. v.10 5-aminolevulinic acid: A potential herbicide/insecticide from microorganisms Sasikala, C.;Ch.V.Ramana;P.R. Rao https://doi.org/10.1021/bp00029a001
  20. Process Biochemistry v.34 Effect of phosphate suply and aeration on poly-β-hydroxybutyrate production in Azotobacter chroococcum Savenkova, L.;Z. Gercberga;Z. Kizhlo;E. Stegantseva https://doi.org/10.1016/S0032-9592(98)00070-3
  21. Seibutsu-kogaku v.73 Inhibitory effect of propionic acid on the 5-aminolevulinic acid (ALA) dehydratase from Rhodobacter sphaeroides and ALA excretion Tanaka, T.;K. Watanabe;S. Nishikawa;K. Sasaki;N. Nishio;S. Nagai
  22. Appl. Environ. Microbiol. v.62 5-aminolevulinate production by Escherichia coli containing the Rhodobacter sphaeroides hemA gene Van der Werf, M.J.;G. Zeikus
  23. Biotechnol. Lett. v.19 H₂and polyhydroxybutyrate, two alternative chemicals from purple non sulfur bacteria Vincenzini, M.;A. Marchini;A. Ena;R.D. Philippis https://doi.org/10.1023/A:1018336209252
  24. Appl. Microbiol. Biotechnol. v.40 Production of a co-polyester of 3-hydroxybutyric acid and 3-hydroxyvaleric acid from succinic acid by Rhodococcus ruber biosynthetic considerations Williams, D.R.;A.J. Anderson;E.A. Dawes;D.F. Ewing https://doi.org/10.1007/BF00173334
  25. J. Ferment. Bioeng. v.80 Effects of amino acid addition on molar fraction of 3-hydroxyvalerate in copolyester of 3-hydroxybutyrate and 3-hydroxyvalerate synthesized by Alcaligenes sp. SH-69 Yoon, J.S.;J.Y. Kim;Y.H. Rhee https://doi.org/10.1016/0922-338X(95)94203-4
  26. Microorganisms and Industry v.22 Effects of amino acid on the biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) in Alcaligengs sp. K-912 Yoon, J.S.;K.Y. Yun;J.Y. Kim;Y.H. Rhee