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4-Hydroxyisoleucine production of recombinant Corynebacterium glutamicum ssp. lactofermentum under optimal corn steep liquor limitation

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Abstract

4-Hydroxyisoleucine (4-HIL) is a nonproteinogenic amino acid that exhibits insulinotropic biological activity. Here, L-isoleucine dioxygenase gene (ido) derived from Bacillus thuringiensis YBT-1520 was cloned and expressed in an L-isoleucine-producing strain, Corynebacterium glutamicum ssp. lactofermentum SN01, in order to directly convert its endogenous L-isoleucine (Ile) into 4-HIL through single-step fermentation. The effects of corn steep liquor limitation as well as ido and truncated idoΔ6 overexpression on 4-HIL production were researched. 4-HIL production by ido-overexpressing strain was improved to 65.44 ± 2.27 mM after fermented for 144 h under corn steep liquor–subsufficient condition, obviously higher than that under corn steep liquor–rich and insufficient conditions. The conversion ratio of Ile to 4-HIL increased to 0.85 mol/mol. In addition, 4-HIL production by ido-overexpressing strain was higher than that by idoΔ6-overexpressing strain, in accord with the relatively higher affinity of Ido as compared to IdoΔ6. This research generated a novel system for 4-HIL de novo biosynthesis and demonstrated corn steep liquor limitation as a useful strategy for improving 4-HIL production in recombinant C. glutamicum ssp. lactofermentum.

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References

  • Alcock NW, Crout DHG, Gregorio MVM, Lee E, Pike G, Samuel CJ (1989) Stereochemistry of the 4-hydroxyisoleucine from Trigonella foenum-graecum. Phytochemistry 28:1835–1841

    Article  CAS  Google Scholar 

  • Asakura Y, Kimura E, Usuda Y, Kawahara Y, Matsui K, Osumi T, Nakamatsu T (2007) Altered metabolic flux due to deletion of odhA causes L-glutamate overproduction in Corynebacterium glutamicum. Appl Environ Microbiol 73:1308–1319. doi:10.1128/AEM. 01867-06

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Blombach B, Schreiner ME, Holatko J, Bartek T, Oldiges M, Eikmanns BJ (2007) L-valine production with pyruvate dehydrogenase complex-deficient Corynebacterium glutamicum. Appl Environ Microbiol 73:2079–2084. doi:10.1128/AEM. 02826-06

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Broca C, Gross R, Petit P, Sauvaire Y, Manteghetti M, Tournier M, Masiello P, Gomis R, Ribes G (1999) 4-Hydroxyisoleucine: experimental evidence of its insulinotropic and antidiabetic properties. Am J Physiol Endocrinol Metab 277:E617–E623

    CAS  Google Scholar 

  • Broca C, Manteghetti M, Gross R, Baissac Y, Jacob M, Petit P, Sauvaire Y, Ribes G (2000) 4-Hydroxyisoleucine: effects of synthetic and natural analogues on insulin secretion. Eur J Pharmacol 390:339–345. doi:10.1016/S0014-2999(00)00030-3

    Article  CAS  PubMed  Google Scholar 

  • Broca C, Breil V, Cruciani-Guglielmacci C, Manteghetti M, Rouault C, Derouet M, Rizkalla S, Pau B, Petit P, Ribes G, Ktorza A, Gross R, Reach G, Taouis M (2004) Insulinotropic agent ID-1101 (4-hydroxyisoleucine) activates insulin signaling in rat. Am J Physiol Endocrinol Metab 287:E463–E471. doi:10.1152/ajpendo.00163.2003

    Article  CAS  PubMed  Google Scholar 

  • Cao Y, Duan ZY, Shi ZP (2014) Effect of biotin on transcription levels of key enzymes and glutamate efflux in glutamate fermentation by Corynebacterium glutamicum. World J Microbiol Biotechnol 30:461–468. doi:10.1007/s11274-013-1468-0

    Article  CAS  PubMed  Google Scholar 

  • Eidi A, Eidi M, Sokhteh M (2007) Effect of fenugreek (Trigonella foenum-graecum L) seeds on serum parameters in normal and streptozotocin-induced diabetic rats. Nutr Res 27:728–733. doi:10.1016/j.nutres.2007.09.006

    Article  CAS  Google Scholar 

  • Fowden L, Pratt HM, Smith A (1973) 4-Hydroxyisoleucine from seed of Trigonella foenum-graecum. Phytochemistry 12:1707–1711

    Article  CAS  Google Scholar 

  • Haeri MR, Limaki HK, White CJ, White KN (2012) Non-insulin dependent anti-diabetic activity of (2S, 3R, 4S) 4-hydroxyisoleucine of fenugreek (Trigonella foenum graecum) in streptozotocin- induced type I diabetic rats. Phytomedicine 19:571–574. doi:10.1016/j.phymed.2012.01.004

    Article  CAS  PubMed  Google Scholar 

  • Hibi M, Kawashima T, Kodera T, Smirnov SV, Sokolov PM, Sugiyama M, Shimizu S, Yokozeki K, Ogawa J (2011) Characterization of Bacillus thuringiensis L-isoleucine dioxygenase for production of useful amino acids. Appl Environ Microbiol 77:6926–6930. doi:10.1128/AEM. 05035-11

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Hirasawa T, Kim J, Shirai T, Furusawa C, Shimizu H (2012) Molecular mechanisms and metabolic engineering of glutamate overproduction in Corynebacterium glutamicum. Subcell Biochem 64:261–281. doi:10.1007/978-94-007-5055-5_13

    Article  CAS  PubMed  Google Scholar 

  • Hu JY, Li YY, Zhang HL, Tan YZ, Wang XY (2014) Construction of a novel expression system for use in Corynebacterium glutamicum. Plasmid 75:18–26. doi:10.1016/j.plasmid.2014.07.005

    Article  CAS  PubMed  Google Scholar 

  • Ikeda M, Nakagawa S (2003) The Corynebacterium glutamicum genome: features and impacts on biotechnological processes. Appl Microbiol Biotechnol 62:99–109. doi:10.1007/s00253-003-1328-1

    Article  CAS  PubMed  Google Scholar 

  • Jaiswal N, Maurya CK, Venkateswarlu K, Sukanya P, Srivastava AK, Narender T, Tamrakar AK (2012) 4-Hydroxyisoleucine stimulates glucose uptake by increasing surface GLUT4 level in skeletal muscle cells via phosphatidylinositol-3-kinase-dependent pathway. Eur J Nutr 51:893–898. doi:10.1007/s00394-012-0374-9

    Article  CAS  PubMed  Google Scholar 

  • Kalinowski J, Bathe B, Bartels D, Bischoff N, Bott M, Burkovski A, Dusch N, Eggeling L, Eikmanns BJ, Gaigalat L, Goesmann A, Hartmann M, Huthmacher K, Kramer R, Linke B, McHardy AC, Meyer F, Mockel B, Pfefferle W, Puhler A, Rey DA, Ruckert C, Rupp O, Sahm H, Wendisch VF, Wiegrabe I, Tauch A (2003) The complete Corynebacterium glutamicum ATCC 13032 genome sequence and its impact on the production of L-aspartate-derived amino acids and vitamins. J Biotechnol 104:5–25. doi:10.1016/S0168-1656(03)00154-8

    Article  CAS  PubMed  Google Scholar 

  • Kawahara Y, Takahashi-Fuke K, Shimizu E, Nakamatsu T, Nakamori S (1997) Relationship between the glutamate production and the activity of 2-oxoglutarate dehydrogenase in Brevibacterium lactofermentum. Biosci Biotechnol Biochem 61:1109–1112

    Article  CAS  PubMed  Google Scholar 

  • Kennerknecht N, Sahm H, Yen MR, Patek M, Saier MH, Eggeling L (2002) Export of L-isoleucine from Corynebacterium glutamicum: a two-gene-encoded member of a new translocator family. J Bacteriol 184:3947–3956. doi:10.1128/JB.184.14.3947-3956.2002

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Kimura E (2003) Metabolic engineering of glutamate production. Adv Biochem Eng Biotechnol 79:37–57

    CAS  PubMed  Google Scholar 

  • Kivero AD, Novikova AE, Smirnov SV (2012) Modification of E. coli central metabolism to optimize the biotransformation of L-isolecucine into 4-hydroxyisoleucine by enzymatic hydroxylation. Appl Biochem Microbiol 48:639–644. doi:10.1134/S0003683812070034

    Article  CAS  Google Scholar 

  • Kodera T, Smirnov SV, Samsonova NN, Kozlov YI, Koyama R, Hibi M, Ogawa J, Yokozeki K, Shimizu S (2009) A novel L-isoleucine hydroxylating enzyme, L-isoleucine dioxygenases from Bacillus thuringiensis, produces (2S,3R,4S)-4-hydroxyisoleucine. Biochem Biophys Res Commun 390:506–510. doi:10.1016/j.bbrc.2009.09.126

    Article  CAS  PubMed  Google Scholar 

  • Lv Y, Liao J, Wu Z, Han S, Lin Y, Zheng S (2012) Genome sequence of Corynebacterium glutamicum ATCC 14067, which provides insight into amino acid biosynthesis in coryneform bacteria. J Bacteriol 194:742–743. doi:10.1128/JB.06514-11

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Maurya CK, Singh R, Jaiswal N, Venkateswarlu K, Narender T, Tamrakar AK (2014) 4-Hydroxyisoleucine ameliorates fatty acid-induced insulin resistance and inflammatory response in skeletal muscle cells. Mol Cell Endocrinol 395:51–60. doi:10.1016/j.mce.2014.07.018

    Article  CAS  PubMed  Google Scholar 

  • Narender T, Puri A, Shweta KT, Saxena R, Bhatia G, Chandra R (2006) 4-hydroxyisoleucine an unusual amino acid as antidyslipidemic and antihyperglycemic agent. Bioorg Med Chem Lett 16:293–296. doi:10.1016/j.bmcl.2005.10.003

    Article  CAS  PubMed  Google Scholar 

  • Ogawa J, Kodera T, Smirnov SV, Hibi M, Samsonova NN, Koyama R, Yamanaka H, Mano J, Kawashima T, Yokozeki K, Shimizu S (2011) A novel L-isoleucine metabolism in Bacillus thuringiensis generating (2S,3R,4S)-4-hydroxyisoleucine, a potential insulinotropic and anti-obesity amino acid. Appl Microbiol Biotechnol 89:1929–1938. doi:10.1007/s00253-010-2983-7

    Article  CAS  PubMed  Google Scholar 

  • Park JH, Lee SY (2010) Fermentative production of branched chain amino acids: a focus on metabolic engineering. Appl Microbiol Biotechnol 85:491–506. doi:10.1007/s00253-009-2307-y

    Article  CAS  PubMed  Google Scholar 

  • Sato H, Orishimo K, Shirai T, Hirasawa T, Nagasawa K, Shimizu H, Wachi M (2008) Distinct roles of two anaplerotic pathways in glutamate production induced by biotin limitation in Corynebacterium glutamicum. J Biosci Bioeng 106:51–58. doi:10.1263/jbb.106.51

    Article  CAS  PubMed  Google Scholar 

  • Sauvaire Y, Petit P, Broca C, Manteghetti M, Baissac Y, Fernandez-Alvarez J, Gross R, Roye M, Leconte A, Gomis R, Ribes G (1998) 4-Hydroxyisoleucine: a novel amino acid potentiator of insulin secretion. Diabetes 47:206–210. doi:10.2337/diabetes.47.2.206

    Article  CAS  PubMed  Google Scholar 

  • Schneider J, Peters-Wendisch P, Stansen KC, Gotker S, Maximow S, Kramer R, Wendisch VF (2012) Characterization of the biotin uptake system encoded by the biotin-inducible bioYMN operon of Corynebacterium glutamicum. BMC Microbiol 12:6. doi:10.1186/1471-2180-12-6

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Shi F, Huan XJ, Wang XY, Ning JF (2012) Overexpression of NAD kinases improves the L-isoleucine biosynthesis in Corynebacterium glutamicum ssp. lactofermentum. Enzym Microbiol Technol 51:73–80. doi:10.1016/j.enzmictec.2012.04.003

    Article  CAS  Google Scholar 

  • Shi F, Li K, Huan XJ, Wang XY (2013) Expression of NAD(H) kinase and glucose-6-phosphate dehydrogenase improve NADPH supply and L-isoleucine biosynthesis in Corynebacterium glutamicum ssp. lactofermentum. Appl Biochem Biotechnol 171:504–521. doi:10.1007/s12010-013-0389-6

    Article  CAS  PubMed  Google Scholar 

  • Singh AB, Tamarkar AK, Narender T, Srivastava AK (2010) Antihyperglycaemic effect of an unusual amino acid (4-hydroxyisoleucine) in C57BL/KsJ-db/db mice. Nat Prod Res 24:258–265. doi:10.1080/14786410902836693

    Article  CAS  PubMed  Google Scholar 

  • Smirnov SV, Kodera T, Samsonova NN, Kotlyarova VA, Rushkevich NY, Kivero AD, Sokolov PM, Hibi M, Ogawa J, Shimizy S (2010) Metabolic engineering of Escherichia coli to produce (2S,3R,4S)-4-hydroxyisoleucine. Appl Microbiol Biotechnol 88:719–726. doi:10.1007/s00253-010-2772-3

    Article  CAS  PubMed  Google Scholar 

  • Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Biotechnology 24:145–149

    Google Scholar 

  • Wu D, He J, Gong Y, Chen D, Zhu X, Qiu N, Sun M, Li M, Yu Z (2011) Proteomic analysis reveals the strategies of Bacillus thuringiensis YBT-1520 for survival under long-term heat stress. Proteomics 11:2580–2591. doi:10.1002/pmic.201000392

    Article  CAS  PubMed  Google Scholar 

  • Yin LH, Shi F, Hu XQ, Chen C, Wang XY (2013) Increasing l-isoleucine production in Corynebacterium glutamicum by overexpressing global regulator Lrp and two-component export system BrnFE. J Appl Microbiol 114:1369–1377. doi:10.1111/jam.12141

    Article  CAS  PubMed  Google Scholar 

  • Yukawa H, Omumasaba CA, Nonaka H, Kos P, Okai N, Suzuki N, Suda M, Tsuge Y, Watanabe J, Ikeda Y, Vertes AA, Inui M (2007) Comparative analysis of the Corynebacterium glutamicum group and complete genome sequence of strain R. Microbiology 153:1042–1058. doi:10.1099/mic. 0.2006/003657-0

    Article  CAS  PubMed  Google Scholar 

  • Zhang D, Liang N, Shi Z, Liu L, Chen J, Du G (2009) Enhancement of α-ketoglutarate production in Torulopsis glabrata: redistribution of carbon flux from pyruvate to α-ketoglutarate. Biotechnol Bioprocess Eng 14:134–139. doi:10.1007/s12257-008-0169-2

    Article  CAS  Google Scholar 

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Acknowledgments

The authors thank the “Program of State Key Laboratory of Food Science and Technology” (contract no. SKLF-ZZB-201405 and SKLF-ZZB-201501) for financial support.

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    Authors and Affiliations

    1. State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, China

      Feng Shi, Tengfei Niu & Huimin Fang

    2. Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, China

      Feng Shi, Tengfei Niu & Huimin Fang

    3. Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, Wuxi, 214122, China

      Feng Shi, Tengfei Niu & Huimin Fang

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    1. Feng Shi
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    Correspondence to Feng Shi.

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    Feng Shi and Tengfei Niu contributed equally to this work.

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    Shi, F., Niu, T. & Fang, H. 4-Hydroxyisoleucine production of recombinant Corynebacterium glutamicum ssp. lactofermentum under optimal corn steep liquor limitation. Appl Microbiol Biotechnol 99, 3851–3863 (2015). https://doi.org/10.1007/s00253-015-6481-9

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