Standardized biosynthesis of flavan-3-ols with effects on pancreatic beta-cell insulin secretion
- 435 Downloads
Flavan-3-ols, such as green tea catechins represent a major group of phenolic compounds with significant medicinal properties. We describe the construction and optimization of Escherichia coli recombinant strains for the production of mono- and dihydroxylated catechins from their flavanone and phenylpropanoid acid precursors. Use of glucose minimal medium, Fe(II), and control of oxygen availability during shake-flask experiments resulted in production yield increases. Additional production improvement resulted from the use of medium rather than high-copy number plasmids and, in the case of mono-hydroxylated compounds, the addition of extracellular cofactors in the culture medium. The established metabolic engineering approach allowed the biosynthesis of natural catechins at high purity for assessing their possible insulinotropic effects in pancreatic β-cell cultures. We demonstrated that (+)-afzelechin and (+)-catechin modulated the secretion of insulin by pancreatic β-cells. These results indicate the potential of applying metabolic engineering approaches for the synthesis of natural and non-natural catechin analogues as drug candidates in diabetes treatments.
KeywordsFlavonoids Catechins Metabolic engineering Insulin Diabetes Flavan-3-ols.
We thank the members of the Koffas and Tzanakakis laboratories for helpful discussions. E.S.T. is supported by a J.D. Watson Award from the New York State Foundation for Science, Technology and Innovation (NYSTAR). M.A.G.K. and E.S.T. acknowledge the support received from the New York State Center of Excellence in Bioinformatics and Life Sciences.
- Ahmad F, Khan MM, Rastogi AK, Chaubey M, Kidwai JR (1991) Effect of (−)-epicatechin on cAMP content, insulin release and conversion of proinsulin to insulin in immature and mature rat islets in vitro. Indian J Exp Biol 29:516–520Google Scholar
- Britsch L, Ruhnau-Brich B, Forkmann G (1992) Molecular cloning, sequence analysis, and in vitro expression of flavanone 3 beta-hydroxylase from Petunia hybrida. J Biol Chem 267:5380–5387Google Scholar
- Deters DW, Racker E, Nelson N, Nelson H (1975) Partial resolution of the enzymes catalyzing photophosphorylation. XV. Approaches to the active site of coupling factor I. J Biol Chem 250:1041–1047Google Scholar
- Ferner RE, Neil HA (1988) Sulphonylureas and hypoglycaemia. Br Med J (Clin Res Ed) 296:949–950Google Scholar
- Kappus H, Koster-Albrecht D, Remmer H (1979) 2-Hydroxyoestradiol and (+)-cyanidanol-3 prevent lipid peroxidation of isolated rat hepatocytes. Arch Toxicol Suppl 59:321–326Google Scholar
- Li G, Min BS, Zheng C, Lee J, Oh SR, Ahn KS, Lee HK (2005) Neuroprotective and free radical scavenging activities of phenolic compounds from Hovenia dulcis. Arch Pharm Res 28:804–809Google Scholar
- Middleton E Jr., Kandaswami C, Theoharides TC (2000) The effects of plant flavonoids on mammalian cells: implications for inflammation, heart disease, and cancer. Pharmacol Rev 52:673–751Google Scholar
- Moroney MA, Alcaraz MJ, Forder RA, Carey F, Hoult JR (1988) Selectivity of neutrophil 5-lipoxygenase and cyclo-oxygenase inhibition by an anti-inflammatory flavonoid glycoside and related aglycone flavonoids. J Pharm Pharmacol 40:787–792Google Scholar
- Negre-Salvayre A, Alomar Y, Troly M, Salvayre R (1991) Ultraviolet-treated lipoproteins as a model system for the study of the biological effects of lipid peroxides on cultured cells. III. The protective effect of antioxidants (probucol, catechin, vitamin E) against the cytotoxicity of oxidized LDL occurs in two different ways. Biochim Biophys Acta 1096:291–300Google Scholar
- Ohigashi H, Minami S, Fukui H, Koshimizu K, Mizutani F, Sugiura A, Tomana T (1982) Flavanols, as plant growth inhibitors from roots of peach, Prunus persica Batsh. cv. “Hakuto”. Agric Biol Chem 46:2555–2561Google Scholar
- Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NYGoogle Scholar
- Tanner GJ, Ashton AR, Arahams S, Watson JM, Larkin PJ, Francki KT (2002) Leucoanthocyanidin reductase and its encoding gene from Desmodium uncinatum and its uses in modifying the pasture qualities of crops. Int Patent Appl WO 2002066625 A1 20020829Google Scholar
- Videla LA, Valenzuela A, Fernandez V, Kriz A (1985) Differential lipid peroxidative response of rat liver and lung tissues to glutathione depletion induced in vivo by diethyl maleate: effect of the antioxidant flavonoid (+)-cyanidanol-3. Biochem Int 10:425–433Google Scholar
- Wolfram S, Raederstorff D, Preller M, Wang Y, Teixeira SR, Riegger C, Weber P (2006) Epigallocatechin gallate supplementation alleviates diabetes in rodents. J Nutr 136:2512–2518Google Scholar