Efficient acetoin production from pyruvate by engineered <i>Halomonas bluephagenesis</i> whole-cell biocatalysis

doi:10.1007/s11705-022-2229-0

Front. Chem. Sci. Eng.

2023, Vol. 17

Issue (4) : 425-436 https://doi.org/10.1007/s11705-022-2229-0

RESEARCH ARTICLE

Efficient acetoin production from pyruvate by engineered Halomonas bluephagenesis whole-cell biocatalysis

Meiyu Zheng^1,², Zhenzhen Cui^1,², Jing Zhang^1,², Jing Fu³, Zhiwen Wang^1,², Tao Chen^1,²(

)

¹. Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300350, China
². Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin 300350, China
³. Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg 41296, Sweden

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Abstract

Acetoin is an important platform chemical, which has a wide range of applications in many industries. Halomonas bluephagenesis, a chassis for next generation of industrial biotechnology, has advantages of fast growth and high tolerance to organic acid salts and alkaline environment. Here, α-acetolactate synthase and α-acetolactate decarboxylase from Bacillus subtilis 168 were co-expressed in H. bluephagenesis to produce acetoin from pyruvate. After reaction condition optimization and further increase of α-acetolactate decarboxylase expression, acetoin production and yield were significantly enhanced to 223.4 mmol·L^–1 and 0.491 mol·mol^–1 from 125.4 mmol·L^–1 and 0.333 mol·mol^–1, respectively. Finally, the highest titer of 974.3 mmol·L^–1 (85.84 g·L^–1) of acetoin was accumulated from 2143.4 mmol·L^–1 (188.6 g·L^–1) of pyruvic acid within 8 h in fed-batch bioconversion under optimal reaction conditions. Moreover, the reusability of the cell catalysis was also tested, and the result illustrated that the whole-cell catalysis obtained 433.3, 440.2, 379.0, 442.8 and 339.4 mmol·L^–1 (38.2, 38.8, 33.4, 39.0 and 29.9 g·L^–1) acetoin in five repeated cycles under the same conditions. This work therefore provided an efficient H. bluephagenesis whole-cell catalysis with a broad development prospect in biosynthesis of acetoin.

Keywords acetoin pyruvate α-acetolactate synthetase α-acetolactate decarboxylase Halomonas bluephagenesis whole-cell biocatalysis

Corresponding Author(s): Tao Chen

Online First Date: 17 January 2023 Issue Date: 24 March 2023

Cite this article:

Meiyu Zheng,Zhenzhen Cui,Jing Zhang, et al. Efficient acetoin production from pyruvate by engineered Halomonas bluephagenesis whole-cell biocatalysis[J]. Front. Chem. Sci. Eng., 2023, 17(4): 425-436.

URL:

https://academic.hep.com.cn/fcse/EN/10.1007/s11705-022-2229-0
https://academic.hep.com.cn/fcse/EN/Y2023/V17/I4/425

Tab.1 Stains and plasmids used in this study

Fig.1 Scheme of acetoin production from pyruvate catalyzed by H. bluephagenesis (PDH, pyruvate dehydrogenase complex; PTA, phosphate acetyltransferase; ACK, acetate kinase).

Fig.2 Acetoin yield, acetoin productivity and maximum acetoin concentration of engineered strains TDZ-1 to TDZ-4 (Error bars represent the standard deviation of triplicate experiments).

Fig.3 The process optimization for efficient acetoin biosynthesis using TDZ-4. (a) The effect of temparature on pyruvate consumption; (b) the effect of temparature on acetoin production; (c) the effect of Mg²⁺ concentration on pyruvate consumption; (d) the effect of Mg²⁺ concentration on acetoin production; (e) the effect of pH on acetoin production; (f) the effect of cell dry weight on acetoin yield, acetoin productivity and maximum acetoin concentration (Error bars represent the standard deviation of triplicate experiments).

Fig.4 The profile of pyruvate consumption and acetoin production in whole-cell biocatalysis using TDZ-4 and TDZ-5. (a) Pyruvate consumption and acetoin production of TDZ-4 and TDZ-5; (b) acetoin yield and productivity of TDZ-4 and TDZ-5 (Error bars represent the standard deviation of triplicate experiments).

Fig.5 The profile of pyruvate consumption, acetoin production and by-products synthesis under different pyruvate concentration. (a) The effect of pyruvate concentration on pyruvate consumption; (b) the effect of pyruvate concentration on acetoin production; (c) the concentrations of by-products in reaction system under different pyruvate concentrations (Error bars represent the standard deviation of triplicate experiments).

Fig.6 The time profile of pH and acetoin production in fed-batch using TDZ-5. (a) pH value of the reaction system; (b) pyruvate consumption and acetoin production in fed-batch bioconversion of TDZ-5 (Error bars represent the standard deviation of triplicate experiments).

Tab.2 Recycling and reutilization of biocatalysis

Fig.7 Acetoin production, acetoin yield and cell dry weight of TDZ-5 in repeated batch bioconversion. The pyruvate addition concentrations were 867.2, 882.6, 765.7, 907.9 and 853.6 mmol·L^–1, respectively. Error bars represent the standard deviation of triplicate experiments.

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