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Opportunities and Challenges of in vitro Synthetic Biosystem for Terpenoids Production

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An Erratum to this article was published on 24 December 2022

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Abstract

Terpenoids are a large variety of natural products with remarkable diverse biological functions, and have a wide range of applications in flavors, pharmaceuticals, biofuels, pigments, and so on. However, limited production of terpenoids from natural resources constrains their use of bulk commodity products. In vivo synthetic biosystem, harnessing living organisms to produce terpenoids, has been broadly used and in-depth reviewed for terpenoids production, which has inherent weaknesses, such as slow reaction rate, low product yield, toxic intermediates, and high separation cost. In vitro synthetic biosystem, harboring numerous enzymes and/or coenzymes assembled into an in vitro enzymatic bioreactions, can obviate part of problems associated with in vivo style. In this review, the general design of in vitro synthetic biosystem is presented with seven supporting examples: mevalonate, isoprene, limonene, pinene, farnesene, amorpha-4,11-diene and taxadiene. The efforts for the large-scale implementation of in vitro synthetic biosystem have been addressed to enzymes engineering, computational modeling and cofactors recycle. The review also discusses the challenges and solutions for the large-scale implementation of in vitro synthetic biosystem around enzymes stability and cofactors recycle. This review may suggest in vitro synthetic biosystems become a realistic option for the production of diverse valuable terpenoids, even expand to other commodity chemicals.

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Abbreviations

ACAT :

acetyl-CoA acetyltransferase

HMGS :

hydroxymethylglutaryl-CoA synthase

HMGR :

hydroxymethylglutaryl-CoA reductase

ackA :

acetate kinase

pta :

phosphate acetyltransferase

G6PDH :

glucose-6-phosphate dehydrogenase

GsPDH :

pyruvate dehydrogenase from Geobacillus stearothermophilus, which is specific for NAD+ to produce acetyl-CoA

LlNoxE :

NADH oxidase from Lactobacillus lactis

GsPyk :

pyruvate kinase from G. stearothermophilus

CoPASI:

a biochemical simulator

Hex:

hexokinase

Pdh :

pyruvate dehydrogenase

Pi :

inorganic phosphate

ck :

choline kinase

ipk :

isopentenyl kinase

idi :

isopentenyl pyrophosphate isomerase

ispA :

farnesyl pyrophosphate synthase

ggpps :

geranylgeranyl pyrophosphate synthase

tds :

taxadiene synthase.

GsPDH/LlNoxE system:

the system recycles free CoA and produce acetyl-CoA continuously.

Plackett-Burman experimental design:

this method is a technique for the optimization of reaction components.

Taguchi method:

this method is an engineering method with low cost and high benefit. For the purpose of experimental design, this method describes a large number of experimental situations to reduce experimental errors and enhance the efficiency and reproducibility of laboratory experiments.

ackA-pta pathway:

this pathway plays part in acetate metabolism.

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Acknowledgements

This work is supported by the Research Project Supported by Shanxi Scholarship Council of China (No. 2020-132), the Jinzhong Key Research and Development Program for Science and Technology (No. Y191021), the Science and Technology Innovation Project of Universities in Shanxi province (No. 2019L0721), and the Basic Research Project of Shanxi University of Traditional Chinese Medicine (No. 2020PY-JC(Y)-05).

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Authors and Affiliations

  1. School of Basic Medical Sciences, Shanxi University of Chinese Medicine, 030619, Jinzhong, Shanxi, China

    Yang Liyang, Gong Qiang & Li Guilan

  2. School of Public Health, Shanxi Medical University, 030001, Taiyuan, Shanxi, China

    Lv Jifang, Zhou Bangyuan & Guo JianQuan

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  1. Yang Liyang
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Liyang, Y., Qiang, G., Jifang, L. et al. Opportunities and Challenges of in vitro Synthetic Biosystem for Terpenoids Production. Biotechnol Bioproc E 27, 697–705 (2022). https://doi.org/10.1007/s12257-022-0100-2

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