In vitro production of n-butanol from glucose

Krutsakorn, Borimas; Honda, Kohsuke; Ye, Xiaoting; Imagawa, Takashi; Bei, Xiaoyu; Okano, Kenji; Ohtake, Hisao

doi:10.1016/j.ymben.2013.09.006

Cited by 91 publications

(113 citation statements)

References 51 publications

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“…13), the inherently low, but Supplementary Table S3. promiscuous, activities of one enzyme that catalyses several substrates results in very low power densities. The use of more than ten enzymes for implementing complex reactions for the production of biocommodities, fine chemicals and pharmaceuticals seems to be not economically prohibitive 25,26,[30][31][32][33] .…”

Section: Discussionmentioning

confidence: 99%

A high-energy-density sugar biobattery based on a synthetic enzymatic pathway

Zhu

Tam²,

Sun³

et al. 2014

Nat Commun

247

194

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High-energy-density, green, safe batteries are highly desirable for meeting the rapidly growing needs of portable electronics. The incomplete oxidation of sugars mediated by one or a few enzymes in enzymatic fuel cells suffers from low energy densities and slow reaction rates. Here we show that nearly 24 electrons per glucose unit of maltodextrin can be produced through a synthetic catabolic pathway that comprises 13 enzymes in an air-breathing enzymatic fuel cell. This enzymatic fuel cell is based on non-immobilized enzymes that exhibit a maximum power output of 0.8 mW cm À 2 and a maximum current density of 6 mA cm À 2 , which are far higher than the values for systems based on immobilized enzymes. Enzymatic fuel cells containing a 15% (wt/v) maltodextrin solution have an energy-storage density of 596 Ah kg À 1 , which is one order of magnitude higher than that of lithium-ion batteries. Sugar-powered biobatteries could serve as next-generation green power sources, particularly for portable electronics.

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Section: Discussionmentioning

confidence: 99%

A high-energy-density sugar biobattery based on a synthetic enzymatic pathway

Zhu

Tam²,

Sun³

et al. 2014

Nat Commun

247

194

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“…The concept was recently advanced with a creative system that generated hydrogen from cellobiose at nearly 100% yields 16 . In another effort, hyper-thermophilic glycolysis enzymes were heterologously expressed, heat purified and assembled to convert glucose to n-butanol in 82% yield 18 . In another study, an elegantly simplified non-phosphorylative Entner-Doudoroff pathway from hyper-thermophilic archaea was constructed to produce ethanol and isobutanol in B55% yields 19 .…”

mentioning

confidence: 99%

“…Thus, the utilization of NAD þ was perfectly balanced allowing multiple passes through the pathway. In other systems, the pathways were also designed to maintain cofactor balance 18 .…”

mentioning

confidence: 99%

A synthetic biochemistry molecular purge valve module that maintains redox balance

2014

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“…The optimum concentrations of enzymes to achieve a desired flux can be experimentally determined by modulating the concentrations of the rate-limiting enzymes [4,5]. Accordingly, we divided the glycerol converting pathway in three parts, namely from glycerol to DHAP, from DHAP to 3-PG, and from 3-PG to lactate, and then adjusted the enzyme concentrations in each partial pathway separately.…”

Section: Lactate Productionmentioning

confidence: 99%

“…A possible solution to this problem is to avoid the use of living microorganisms and to construct an in vitro artificial metabolic pathway in which only a limited number of enzymes are involved. Until now, a variety of in vitro synthetic pathways have been designed and constructed for the production of alcohols [3,4], organic acids [5,6], carbohydrates [7], hydrogen [8,9], bioplastic [10], and even electricity [11]. Particularly, employment of enzymes derived from thermophiles and hyperthermophiles enables the simple preparation of catalytic modules with excellent selectivity and thermal stability [5,12].…”

Section: Introductionmentioning

confidence: 99%

In vitro conversion of glycerol to lactate with thermophilic enzymes

Jaturapaktrarak

Napathorn

Cheng

et al. 2014

Bioresour. Bioprocess.

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Background: In vitro reconstitution of an artificial metabolic pathway has emerged as an alternative approach to conventional in vivo fermentation-based bioproduction. Particularly, employment of thermophilic and hyperthermophilic enzymes enables us a simple preparation of highly stable and selective biocatalytic modules and the construction of in vitro metabolic pathways with an excellent operational stability. In this study, we designed and constructed an artificial in vitro metabolic pathway consisting of nine (hyper)thermophilic enzymes and applied it to the conversion of glycerol to lactate. We also assessed the compatibility of the in vitro bioconversion system with methanol, which is a major impurity in crude glycerol released from biodiesel production processes. Results: The in vitro artificial pathway was designed to balance the intrapathway consumption and regeneration of energy and redox cofactors. All enzymes involved in the in vitro pathway exhibited an acceptable level of stability at high temperature (60°C), and their stability was not markedly affected by the co-existing of up to 100 mM methanol. The one-pot conversion of glycerol to lactate through the in vitro pathway could be achieved in an almost stoichiometric manner, and 14.7 mM lactate could be produced in 7 h. Furthermore, the in vitro bioconversion system exerted almost identical performance in the presence of methanol. Conclusions: Many thermophilic enzymes exhibit higher stability not only at high temperatures but also in the presence of denaturants such as detergents and organic solvents than their mesophilic counterparts. In this study, compatibilities of thermophilic enzymes with methanol were demonstrated, indicating the potential applicability of in vitro bioconversion systems with thermophilic enzymes in the conversion of crude glycerol to value-added chemicals.

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In vitro production of n-butanol from glucose

Cited by 91 publications

References 51 publications

A high-energy-density sugar biobattery based on a synthetic enzymatic pathway

A high-energy-density sugar biobattery based on a synthetic enzymatic pathway

A synthetic biochemistry molecular purge valve module that maintains redox balance

In vitro conversion of glycerol to lactate with thermophilic enzymes

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