scholarly journals Engineering Corynebacterium glutamicum for the Efficient Production of 3-Hydroxypropionic Acid from a Mixture of Glucose and Acetate via the Malonyl-CoA Pathway

Catalysts ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 203
Author(s):  
Zhishuai Chang ◽  
Wei Dai ◽  
Yufeng Mao ◽  
Zhenzhen Cui ◽  
Zhiwen Wang ◽  
...  

3-Hydroxypropionic acid (3-HP) has been recognized as one of the top value-added building block chemicals, due to its numerous potential applications. Over the past decade, biosynthesis of 3-HP via the malonyl-CoA pathway has been increasingly favored because it is balanced in terms of ATP and reducing equivalents, does not require the addition of costly coenzymes, and can utilize renewable lignocellulosic biomass. In this study, gene mcr encoding malonyl-CoA reductase from Chloroflexus aurantiacus was introduced into Corynebacterium glutamicum ATCC13032 to construct the strain Cgz1, which accumulated 0.30 g/L 3-HP. Gene ldhA encoding lactate dehydrogenase was subsequently deleted to eliminate lactate accumulation, but this decreased 3-HP production and greatly increased acetate accumulation. Then, different acetate utilization genes were overexpressed to reuse the acetate, and the best candidate Cgz5 expressing endogenous gene pta could effectively reduce the acetate accumulation and produced 0.68 g/L 3-HP. To enhance the supply of the precursor acetyl-CoA, acetate was used as an ancillary carbon source to improve the 3-HP production, and 1.33 g/L 3-HP could be produced from a mixture of glucose and acetate, with a 2.06-fold higher yield than from glucose alone. Finally, to inhibit the major 3-HP competing pathway-fatty acid synthesis, 10 μM cerulenin was added and strain Cgz5 produced 3.77 g/L 3-HP from 15.47 g/L glucose and 4.68 g/L acetate with a yield of 187 mg/g substrate in 48 h, which was 12.57-fold higher than that of Cgz1. To our best knowledge, this is the first report on engineering C. glutamicum to produce 3-HP via the malonyl-CoA pathway. The results indicate that the innocuous biosafety level I microorganism C. glutamicum is a potential industrial 3-HP producer.

Author(s):  
Zhishuai Chang ◽  
Wei Dai ◽  
Yufeng Mao ◽  
Zhenzhen Cui ◽  
Zhidan Zhang ◽  
...  

Acetate is an economical and environmental-friendly alternative carbon source. Herein, the potential of harnessing Corynebacterium glutamicum as a host to produce 3-hydroxypropionic acid (3-HP) from acetate was explored. First, the expression level of malonyl-CoA reductase from Chloroflexus aurantiacus was optimized through several strategies, strain Cgz2/sod-N-C* showed an MCR enzyme activity of 63 nmol/mg/min and a 3-HP titer of 0.66 g/L in flasks. Next, the expression of citrate synthase in Cgz2/sod-N-C* was weakened to reduce the acetyl-CoA consumption in the TCA cycle, and the resulting strain Cgz12/sod-N-C* produced 2.39 g/L 3-HP from 9.32 g/L acetate. However, the subsequent deregulation of the expression of acetyl-CoA carboxylase genes in Cgz12/sod-N-C* resulted in an increased accumulation of intracellular fatty acids, instead of 3-HP. Accordingly, cerulenin was used to inhibit fatty acid synthesis in Cgz14/sod-N-C*, and its 3-HP titer was further increased to 4.26 g/L, with a yield of 0.50 g 3-HP/g-acetate. Finally, the engineered strain accumulated 17.1 g/L 3-HP in a bioreactor without cerulenin addition, representing the highest titer achieved using acetate as substrate. The results demonstrated that Corynebacterium glutamicum is a promising host for 3-HP production from acetate.


Catalysts ◽  
2018 ◽  
Vol 8 (11) ◽  
pp. 525 ◽  
Author(s):  
Ji Lee ◽  
Sanghak Cha ◽  
Chae Kang ◽  
Geon Lee ◽  
Hyun Lim ◽  
...  

Acetate, which is an abundant carbon source, is a potential feedstock for microbial processes that produce diverse value-added chemicals. In this study, we produced 3-hydroxypropionic acid (3-HP) from acetate with engineered Escherichia coli. For the efficient conversion of acetate to 3-HP, we initially introduced heterologous mcr (encoding malonyl-CoA reductase) from Chloroflexus aurantiacus. Then, the acetate assimilating pathway and glyoxylate shunt pathway were activated by overexpressing acs (encoding acetyl-CoA synthetase) and deleting iclR (encoding the glyoxylate shunt pathway repressor). Because a key precursor malonyl-CoA is also consumed for fatty acid synthesis, we decreased carbon flux to fatty acid synthesis by adding cerulenin. Subsequently, we found that inhibiting fatty acid synthesis dramatically improved 3-HP production (3.00 g/L of 3-HP from 8.98 g/L of acetate). The results indicated that acetate can be used as a promising carbon source for microbial processes and that 3-HP can be produced from acetate with a high yield (44.6% of the theoretical maximum yield).


2021 ◽  
Vol 10 (1) ◽  
pp. 264-281
Author(s):  
Ao Li ◽  
Dezhong Xu ◽  
Lu Luo ◽  
Yalan Zhou ◽  
Wen Yan ◽  
...  

Abstract The rapid economic growth and environmental concerns have led to high demands on paper and paper-based products in terms of variety, quantity, quality, and specialty. Enhancement and functionalization with additives are constantly required. Moving away from traditional petroleum-based additives, researchers have attempted to use “green” nanoadditives by introducing renewable environmentally friendly nanocellulose. This article studies the functions of nanocellulose as bio-additives (enhancer, retention and filtration reagent, and coating aid) in paper and paper products, and overviews the research development of nanocellulose-based additives and their applications in the paper industry for both efficient production and paper functionalization. The review shows that (1) a variety of nanocellulose-based bioadditives have been reported for various applications in paper and paper-based products, while commercially viable developments are to be advanced; (2) nanocellulose was mostly formulated with other polymer and particles as additives to achieve their synergistic effects; (3) major interests have concentrated on the nanocellulose in the specialty papers as representing more value added products and in the efficient utilization of recycled fibers, which remains most attractive and promising for future development. This report shall provide most useful database information for researchers and industries for paper recycling and enhancement, and paper-based products innovation and application.


Fermentation ◽  
2021 ◽  
Vol 7 (4) ◽  
pp. 220
Author(s):  
Wubliker Dessie ◽  
Zongcheng Wang ◽  
Xiaofang Luo ◽  
Meifeng Wang ◽  
Zuodong Qin

Succinic acid (SA) is one of the top candidate value-added chemicals that can be produced from biomass via microbial fermentation. A considerable number of cell factories have been proposed in the past two decades as native as well as non-native SA producers. Actinobacillus succinogenes is among the best and earliest known natural SA producers. However, its industrial application has not yet been realized due to various underlying challenges. Previous studies revealed that the optimization of environmental conditions alone could not entirely resolve these critical problems. On the other hand, microbial in silico metabolic modeling approaches have lately been the center of attention and have been applied for the efficient production of valuable commodities including SA. Then again, literature survey results indicated the absence of up-to-date reviews assessing this issue, specifically concerning SA production. Hence, this review was designed to discuss accomplishments and future perspectives of in silico studies on the metabolic capabilities of SA producers. Herein, research progress on SA and A. succinogenes, pathways involved in SA production, metabolic models of SA-producing microorganisms, and status, limitations and prospects on in silico studies of A. succinogenes were elaborated. All in all, this review is believed to provide insights to understand the current scenario and to develop efficient mathematical models for designing robust SA-producing microbial strains.


Catalysts ◽  
2018 ◽  
Vol 8 (11) ◽  
pp. 561 ◽  
Author(s):  
Kei-Anne Baritugo ◽  
Hee Taek Kim ◽  
Mi Na Rhie ◽  
Seo Young Jo ◽  
Tae Uk Khang ◽  
...  

Corynebacterium glutamicum is an industrial strain used for the production of valuable chemicals such as L-lysine and L-glutamate. Although C. glutamicum has various industrial applications, a limited number of tunable systems are available to engineer it for efficient production of platform chemicals. Therefore, in this study, we developed a novel tunable promoter system based on repeats of the Vitreoscilla hemoglobin promoter (Pvgb). Tunable expression of green fluorescent protein (GFP) was investigated under one, four, and eight repeats of Pvgb (Pvgb, Pvgb4, and Pvgb8). The intensity of fluorescence in recombinant C. glutamicum strains increased as the number of Pvgb increased from single to eight (Pvgb8) repeats. Furthermore, we demonstrated the application of the new Pvgb promoter-based vector system as a platform for metabolic engineering of C. glutamicum by investigating 5-aminovaleric acid (5-AVA) and gamma-aminobutyric acid (GABA) production in several C. glutamicum strains. The profile of 5-AVA and GABA production by the recombinant strains were evaluated to investigate the tunable expression of key enzymes such as DavBA and GadBmut. We observed that 5-AVA and GABA production by the recombinant strains increased as the number of Pvgb used for the expression of key proteins increased. The recombinant C. glutamicum strain expressing DavBA could produce higher amounts of 5-AVA under the control of Pvgb8 (3.69 ± 0.07 g/L) than the one under the control of Pvgb (3.43 ± 0.10 g/L). The average gamma-aminobutyric acid production also increased in all the tested strains as the number of Pvgb used for GadBmut expression increased from single (4.81–5.31 g/L) to eight repeats (4.94–5.58 g/L).


Author(s):  
Amrut Rao ◽  
Ravindra Pathak ◽  
Ashraf Mahmud Rayed

Ethiopia, India and Bangladesh are raising economic power, but have not yet integrated very much with the global economy and still have not achieved their potential in context of technology, globalization, and international competitiveness like developed countries. These countries have much strength, but at the same time , are facing many challenges in the increasingly competitive and fast changing global economy. The main key strengths of these courtiers are their large domestic market, young and growing population, a strong private sector with experience in market institutions, and a well developed legal and financial system. In today’s environment of global competition, technological development and innovation; companies, especially manufacturing, are forced to reconfigure their manufacturing and management processes. Industry 4.0 and intelligent manufacturing are part of a transformation, in which manufacturing and information technologies have been integrated to create innovative systems of manufacturing, management and ways of doing business. This system allows optimizing manufacturing, to achieve greater flexibility, efficient production processes and generate a value added proposal for their customers, as well as to provide a timely response to their market needs. The objective of this work is to explore the Industry 4.0, smart manufacturing, environment requirement and relation of innovation in perspective of developing countries.


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