Production of Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) by Recombinant Pseudomonas stutzeri 1317 from Unrelated Carbon Sources

doi:10.1016/S1004-9541(13)60549-2

Chinese Journal of Chemical Engineering ›› 2013, Vol. 21 ›› Issue (9): 1057-1061.DOI: 10.1016/S1004-9541(13)60549-2

Production of Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) by Recombinant Pseudomonas stutzeri 1317 from Unrelated Carbon Sources

魏晓星^1,2, 刘峰^2,3, 简嘉², 王瑞妍², 陈国强^2,4

1 Department of Basic Medicine, Medical College of Qinghai University, Xining 810016, China;
2 MOE Key Lab of Bioinformatics and Systems Biology, Department of Biological Science and Biotechnology, School of Life Sciences, Tsinghua-Peking Center for Life Sciences, Tsinghua University, Beijing 100084, China;
3 Tuspark Kunshan Biotech Science Park Development Center, 4/F, Science & Technology Plaza, Tsinghua Science Park, Kunshan, Jiangsu 215347, China;
4 Center for Nano and Micro Mechanics, Tsinghua University, Beijing 100084, China

收稿日期:2012-09-27 修回日期:2013-02-28 出版日期:2013-09-28 发布日期:2013-10-01
通讯作者: CHEN Guoqiang
基金资助:
Supported by the National Natural Science Foundation of China (31260015), Natural Science Foundation of Qinghai Province (2012-Z-919Q), the Extramural Project from State Key Laboratory for Agrobiotechnology (2012SKLAB06-5) and the Research Funds for Young Project of Qinghai University (2011-QYY-1)

Production of Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) by Recombinant Pseudomonas stutzeri 1317 from Unrelated Carbon Sources

WEI Xiaoxing^1,2, LIU Feng^2,3, JIAN Jia², WANG Ruiyan², CHEN Guoqiang^2,4

1 Department of Basic Medicine, Medical College of Qinghai University, Xining 810016, China;
2 MOE Key Lab of Bioinformatics and Systems Biology, Department of Biological Science and Biotechnology, School of Life Sciences, Tsinghua-Peking Center for Life Sciences, Tsinghua University, Beijing 100084, China;
3 Tuspark Kunshan Biotech Science Park Development Center, 4/F, Science & Technology Plaza, Tsinghua Science Park, Kunshan, Jiangsu 215347, China;
4 Center for Nano and Micro Mechanics, Tsinghua University, Beijing 100084, China

Received:2012-09-27 Revised:2013-02-28 Online:2013-09-28 Published:2013-10-01
Supported by:
Supported by the National Natural Science Foundation of China (31260015), Natural Science Foundation of Qinghai Province (2012-Z-919Q), the Extramural Project from State Key Laboratory for Agrobiotechnology (2012SKLAB06-5) and the Research Funds for Young Project of Qinghai University (2011-QYY-1)

摘要/Abstract

摘要： Synthetic biology promises to simplify the construction of metabolic pathways by assembling the detached modules of the whole pathway. This gives new approaches for the microbial production of industrial products such as polyhydroxyalkanoates (PHA). In this study, to produce poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) by Pseudomonas stutzeri 1317 from unrelated carbon sources such as glucose, the phaC1-phaZ-phaC2 operon of P. stutzeri 1317 was knocked out to generate the PHA deficient mutant P. stutzeri 1317LF. Then three modules containing phaC_AhA_ReB_Re, phaC_AhB_ReG_Pp and phaC_AhP_Ah were introduced into P. stutzeri 1317LF separately. The shake flask results indicated that the precursor supply and PHA synthase activity were the vital factors for the PHBHHx accumulation of P. stutzeri 1317LF. Furthermore, the PHBHHx accumulation of the recombinants from different carbon resources were performed. The highest PHBHHx content was 23.7%(by mass) with 58.6%(by mole) 3HB fraction. These results provide basis for further improving the PHBHHx accumulation of P. stutzeri from unrelated carbon sources.

关键词: Pseudomonas stutzeri 1317, PHBHHx, unrelated carbon sources

Abstract: Synthetic biology promises to simplify the construction of metabolic pathways by assembling the detached modules of the whole pathway. This gives new approaches for the microbial production of industrial products such as polyhydroxyalkanoates (PHA). In this study, to produce poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) by Pseudomonas stutzeri 1317 from unrelated carbon sources such as glucose, the phaC1-phaZ-phaC2 operon of P. stutzeri 1317 was knocked out to generate the PHA deficient mutant P. stutzeri 1317LF. Then three modules containing phaC_AhA_ReB_Re, phaC_AhB_ReG_Pp and phaC_AhP_Ah were introduced into P. stutzeri 1317LF separately. The shake flask results indicated that the precursor supply and PHA synthase activity were the vital factors for the PHBHHx accumulation of P. stutzeri 1317LF. Furthermore, the PHBHHx accumulation of the recombinants from different carbon resources were performed. The highest PHBHHx content was 23.7%(by mass) with 58.6%(by mole) 3HB fraction. These results provide basis for further improving the PHBHHx accumulation of P. stutzeri from unrelated carbon sources.

Key words: Pseudomonas stutzeri 1317, PHBHHx, unrelated carbon sources

魏晓星, 刘峰, 简嘉, 王瑞妍, 陈国强. Production of Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) by Recombinant Pseudomonas stutzeri 1317 from Unrelated Carbon Sources[J]. Chinese Journal of Chemical Engineering, 2013, 21(9): 1057-1061.

WEI Xiaoxing, LIU Feng, JIAN Jia, WANG Ruiyan, CHEN Guoqiang, . Production of Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) by Recombinant Pseudomonas stutzeri 1317 from Unrelated Carbon Sources[J]. Chin.J.Chem.Eng., 2013, 21(9): 1057-1061.

参考文献

1 Chen, G.Q., “A microbial polyhydroalkanoates (PHA) based bio- and materials industry”, Chem. Soc. Rev., 38, 2434-2446 (2009).

2 Dowes, E.A., Senior, P.J., “The role and regulation of energy reserve polymers in microorganisms”, Adv. Microb. Physiol., 10, 135-266 (1973).

3 Chen, G.Q., Luo, R.C., Xu, J., Wu, Q., Microbial PHA Based Eco-Material Industry, Chemical Industry Press, Beijing, 1-21 (2008). (in Chinese)

4 Chen, G.Q., Wu, Q., “The application of polyhydroxyalkanoates as tissue engineering materials”, Biomaterials, 26, 6565-6578 (2005).

5 Li, Z.J., Wei, X.X., Chen, G.Q., “Microbial cell factories for production of polyhydroxyalkanoates”, Chin. J. Biotechnol., 26, 1426-1435 (2010).

6 Steinbüchel, A., “Perspectives for biotechnological production and utilization of biopolymers: metabolic engineering of polyhydroxyalkanoate biosynthesis pathways as a successful example”, Macromol. Biosci., 1, 1-24 (2001).

7 Wu, Q., Wang, Y., Chen, G.Q., “Medical application of microbial biopolyesters polyhydroxyalkanoates”, Artif. Cells Blood Substit. Immobil. Biotechnol., 37, 1-12 (2009).

8 Liu, F., Jian, J., Shen, X.W., Chung, A., Chen, J.C., Chen, G.Q., “Metabolic engineering of Aeromonas hydrophila 4AK4 for production of copolymers of 3-hydroxybutyrate and medium-chain-length 3-hydroxyalkanoate”, Bioresour. Technol., 102, 8123-8129 (2011).

9 Lu, X.Y., Zhang, W.J., Jian, J., Wu, Q., Chen, G.Q., “Molecular cloning and functional analysis of two polyhydroxyalkanoate synthases from two strains of Aeromonas hydrophila spp.”, FEMS Microbiol. Lett., 243, 149-155 (2005).

10 Chen, G.Q., Zhang, G., Park, S.J., Lee, S.Y., “Industrial scale production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)”, Appl. Microbiol. Biotechnol., 57, 50-55 (2001).

11 He, W.N., Tian, W.D., Zhang, G., Chen, G.Q., Zhang, Z.M., “Production of novel polyhydroxyalkanoates by Pseudomonas stutzeri 1317 from glucose and soybean oil”, FEMS. Microbiol. Lett., 169, 45-49 (1998).

12 Chen, J.Y., Liu, T., Zheng, Z. Chen, J.C., Chen, G.Q., “Polyhydroxyalkanoate synthases phaC1 and phaC2 from Pseudomonas stutzeri 1317 had different substrate specificities”, FEMS. Microbiol. Lett., 234, 231-237 (2004).

13 Chen, G.Q., Xu, J., Wu, Q., Zhang, Z.M. Ho, K.P., “Synthesis of copolyesters consisting of medium-chain-length β-hydroxyalkanoates by Pseudomonas stutzeri 1317”. React. Funct. Poly., 48, 107-112 (2001).

14 Benner, S.A., Sismour, A.M., “Synthetic biology”, Nat. Rev. Genet., 6, 533-543 (2005).

15 Schäfer, A., Tauch, A., Jäger, W., Kalinowski, J., Thierbach, G., Pühler, A., “Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum”, Gene., 145, 69-73 (1994).

16 Chen, J.Y., Song, G., Chen, G.Q., “A lower specificity phaC2 synthase from Pseudomonas stutzeri catalyses the production of copolyesters consisting of short-chain-length and medium-chain-length 3-hydroxyalkanoates”, Anton Leeuw. Int. J. G., 89, 157-167 (2006).

17 Cai, L., Yuan, M.Q., Liu, F., Chen, G.Q., “Enhanced production of medium-chain-length polyhydroxyalkanoates (PHA) by PHA depolymerase knockout mutant of Pseudomonas putida KT2442”, Bioresour. Technol., 100, 2265-2270 (2009).

18 Chung, A., Liu, Q., Ouyang, S.P., Wu, Q., Chen, G.Q., “Microbial production of 3-hydroxydodecanoic acid by pha operon and fadBA knockout mutant of Pseudomonas putida KT2442 harboring tesB gene”, Appl. Microbiol. Biotechnol., 83, 513-519 (2009).

19 Wei, X.X., Shi, Z.Y., Yuan, M.Q., Chen, G.Q., “Effect of anaerobic promoters on the microaerobic production of polyhydroxybutyrate (PHB) in recombinant Escherichia coli”, Appl. Microbiol. Biotechnol., 82, 703-712 (2009).

20 Yan, Y.L., Yang, J., Dou, Y.T., Chen, M., Ping, S.Z., Peng, J.P., Zhang, W., Yao, Z.Y., Li, H.Q., Liu, W., He, S., Geng, L.Z., Zhang, X.B., Yang, F., Yu, H.Y., Zhan, Y.H., Li, D.H., Lin, Z.L., Wang, Y.P., Elmerich, C., Lin, M., Jin, Q., “Nitrogen fixation island and rhizosphere competence traits in the genome of root-associated Pseudomonas stutzeri A1501”, Proc. Natl. Acad. Sci. USA, 105, 7564-7569 (2008).

21 Rehm, B.H.A., Kruger, N., Steinbüchel, A., “A new metabolic link between fatty acid de novo synthesis and polyhydroxyalkanoic acid synthesis: the phaG gene from Pseudomonas putida KT2440 encodes a 3-hydroxyacyl-acyl carrier protein coenzyme A transferase”, J. Biol. Chem., 273, 24044-24051 (1998).

22 Fukui, T., Kichise, T., Iwata, T., Doi, Y., “Characterization of 13 kDa granule-associated protein in Aeromonas caviae and biosynthesis of polyhydroxy-alkanoates with altered molar composition by recombinant bacteria”, Biomacromolecules, 2, 148-153 (2001).

23 Simon, R., Priefer, U., Pühler, A., “A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in gram negative bacteria”, Nat. Biotechnol., 1, 784-791 (1983).

24 Qiu, Y.Z., Han, J., Guo, J.J., Chen, G.Q., “Production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) from gluconate and glucose by recombinant Aeromonas hydrophila and Pseudomonas putida”, Biotechnol. Lett., 27, 1381-1386 (2005).

25 Lu, X.Y., Wu, Q., Zhang, W.J., Jian, J., Chen, G.Q., “Studies on synthesis of polyhydroxyalkanoate consisting of 3-hydroxyvalerate by Aeromonas hydrophila”, Chin. J. Biotechnol., 20, 779-783 (2004).

Production of Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) by Recombinant Pseudomonas stutzeri 1317 from Unrelated Carbon Sources

Production of Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) by Recombinant Pseudomonas stutzeri 1317 from Unrelated Carbon Sources

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