An in vitro synthetic biology platform for the industrial biomanufacturing of myo‐inositol from starch

Chun You; Ting Shi; Pingping Han; Xigui Zhou; Yi‐Heng Percival Zhang

doi:10.1002/bit.26314

journal article May 08, 2017

An in vitro synthetic biology platform for the industrial biomanufacturing of myo‐inositol from starch

Chun You

Ting Shi

Pingping Han

Xigui Zhou Yi‐Heng Percival Zhang

Biotechnology and Bioengineering Vol. 114 No. 8 pp. 1855-1864 · Wiley

View at Publisher Save 10.1002/bit.26314

Abstract

ABSTRACTMyo‐Inositol (vitamin B8) is widely used in the drug, cosmetic, and food & feed industries. Here, we present an in vitro non‐fermentative enzymatic pathway that converts starch to inositol in one vessel. This in vitro pathway is comprised of four enzymes that operate without ATP or NAD+ supplementation. All enzyme BioBricks are carefully selected from hyperthermophilic microorganisms, that is, alpha‐glucan phosphorylase from Thermotoga maritima, phosphoglucomutase from Thermococcus kodakarensis, inositol 1‐phosphate synthase from Archaeoglobus fulgidus, and inositol monophosphatase from T. maritima. They were expressed efficiently in high‐density fermentation of Escherichia coli BL21(DE3) and easily purified by heat treatment. The four‐enzyme pathway supplemented with two other hyperthermophilic enzymes (i.e., 4‐α‐glucanotransferase from Thermococcus litoralis and isoamylase from Sulfolobus tokodaii) converts branched or linear starch to inositol, accomplishing a very high product yield of 98.9 ± 1.8% wt./wt. This in vitro (aeration‐free) biomanufacturing has been successfully operated on 20,000‐L reactors. Less costly inositol would be widely added in heath food, low‐end soft drink, and animal feed, and may be converted to other value‐added biochemicals (e.g., glucarate). This biochemical is the first product manufactured by the in vitro synthetic biology platform on an industrial scale. Biotechnol. Bioeng. 2017;114: 1855–1864. © 2017 Wiley Periodicals, Inc.

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Details

Published: May 08, 2017
Vol/Issue: 114(8)
Pages: 1855-1864
License: View

Authors

C

Chun You