Fermentation of biodiesel-derived glycerol by Bacillus amyloliquefaciens: effects of co-substrates on 2,3-butanediol production

Yang, Taowei; Rao, Zhiming; Zhang, Xian; Xu, Meijuan; Xu, Zhenghong; Yang, Shang‐Tian

doi:10.1007/s00253-013-5048-x

Cited by 49 publications

(25 citation statements)

References 26 publications

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“…Firstly, fed-batch fermentation was performed under the combined feeding strategy (initial addition of beet molasses and later co-feeding with glycerol and molasses) [ 11 ]. The representative time courses of fed-batch fermentation by B. amyloliquefaciens GAR are presented in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…For 2,3-BD production, B. amyloliquefaciens was inoculated into 10 mL LB medium added with 40 g/L glycerol and cultivated overnight with agitation (180 rpm, rotary shaker) at 37 °C. After 12 h, 2.5 mL of the seed culture (OD 600 = 5.0–6.0) was inoculated into fermentation medium (crude glycerol (80 g/L), beet molasses (15 g/L) (Addition of beet molasses could enhance glycerol assimilation [ 11 ] ), corn steep liquor (30 g/L), soybean meal (20 g/L), ammonium citrate (5 g/L), K 2 HPO 4 (3 g/L), MgSO 4 ·7H 2 O (0.3 g/L), FeSO 4 ·7H 2 O (0.05 g/L), pH6.5). Waste glycerol comprised of 88 % (w/w) glycerol, 6–9 % (w/w) water, 4–6 % (w/w) ash, 4 % (w/w) chlorides, and 0.2 % (w/w) methanol.…”

Section: Methodsmentioning

confidence: 99%

“…Previously, we reported that B. amyloliquefaciens readily produces 2,3-BD from biodiesel-derived glycerol in the presence of beet molasses as a co-substrate [ 11 ]. In fed-batch fermentation, 2,3-BD production (83.3 g/L) from waste glycerol reached the highest level reported to date, but fermentation was accompanied by undesirably large production of acetoin, lactate, acetate, and succinate.…”

Section: Introductionmentioning

confidence: 99%

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Enhanced 2,3-butanediol production from biodiesel-derived glycerol by engineering of cofactor regeneration and manipulating carbon flux in Bacillus amyloliquefaciens

et al. 2015

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BackgroundBacillus amyloliquefaciens B10-127 exhibited an excellent ability for industrial-scale microbial fermentation of 2,3-butanediol (2,3-BD) from biodiesel-derived glycerol. However, the accumulation of by-products (acetoin, acetoin, lactate and succinate) and the 2,3-BD yield remains prohibitively low for commercial production.ResultsSeveral strategies were developed to manipulate the carbon flux to 2,3-BD branch in a designed B. amyloliquefaciens. Firstly, extra copies of NADH/NAD+ regeneration system were introduced into B. amyloliquefaciens by co-overproduction of glycerol dehydrogenase and acetoin reductase, which resulting in improvement of 2,3-BD production and suppression of by-products accumulation. Subsequently, the transcriptional regulator ALsR under the control of a moderate promoter PbdhA was introduced into B. amyloliquefaciens, which increased carbon flux to 2,3-BD branch. Finally, a three-stage dissolved oxygen control strategy were proposed based on analysis of the characteristic of 2,3-BD fermentation, and a two-stage pH control strategy were proposed based on different pH preferences of ACR for reduction and oxidation. Following these strategies, a high titer (102.3 g/L), yield (0.44 g/g), and productivity (1.16 g/L/h) of 2,3-BD were achieved.ConclusionsTo our knowledge, this is the highest reported 2,3-BD production using biodiesel-derived glycerol as substrate, and this designed B. amyloliquefaciens should be an excellent candidate for producing 2,3-BD on an industrial scale.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Enhanced 2,3-butanediol production from biodiesel-derived glycerol by engineering of cofactor regeneration and manipulating carbon flux in Bacillus amyloliquefaciens

et al. 2015

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“…This inhibition probably resulted from a decrease in water activity that affects the metabolic rates [32]. Fed-batch fermentation can help to avoid the influence of high concentration substrate limitation and inhibition by controlling the substrate concentration below the toxic level [33,34]. In this study, the glutamic acid production was also improved under fed-batch SSF.…”

Section: Discussionmentioning

confidence: 80%

Two-step production of gamma-aminobutyric acid from cassava powder using Corynebacterium glutamicum and Lactobacillus plantarum

Yang

Rao

Kimani

et al. 2015

Journal of Industrial Microbiology and Biotechnology

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Production of gamma-aminobutyric acid (GABA) from crop biomass such as cassava in high concentration is desirable, but difficult to achieve. A safe biotechnological route was investigated to produce GABA from cassava powder by C. glutamicum G01 and L. plantarum GB01-21. Liquefied cassava powder was first transformed to glutamic acid by simultaneous saccharification and fermentation with C. glutamicum G01, followed by biotransformation of glutamic acid to GABA with resting cells of L. plantarum GB01-21 in the reaction medium. After optimizing the reaction conditions, the maximum concentration of GABA reached 80.5 g/L with a GABA productivity of 2.68 g/L/h. This is the highest yield ever reported of GABA production from cassava-derived glucose. The bioprocess provides the added advantage of employing nonpathogenic microorganisms, C. glutamicum and L. plantarum, in microbial production of GABA from cassava biomass, which can be used in the food and pharmaceutical industries.

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“…Adding sugars, mannitol, or hemicellulose hydrolysate as a cosubstrate to glycerol can significantly increase the yield of 1,3-propanediol production (31)(32)(33). Beet molasses added as a cosubstrate promoted 2,3-butanediol production from biodiesel derived glycerol (34). Although these cosubstrates contributed to the production of cell biomass and regeneration of reducing power, the cosubstrates themselves were not converted into products and represent an additional processing cost.…”

Section: Discussionmentioning

confidence: 99%

Homoethanol Production from Glycerol and Gluconate Using Recombinant Klebsiella oxytoca Strains

Tao

Wang

Walters

et al. 2019

Appl Environ Microbiol

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Gluconic acid, an oxidized cellulose degradation product, could be produced from cellulosic biomass. Glycerol is an inexpensive and renewable resource for fuels and chemicals production and is available as a byproduct of biodiesel production. Gluconate is a more oxidized substrate than glucose, whereas glycerol is a more reduced substrate than glucose. Although the production of homoethanol from glucose can be achieved, the conversion of gluconate to ethanol is accompanied by the production of oxidized byproduct such as acetate, and reduced byproducts such as 1,3-propanediol are produced, along with ethanol, when glycerol is used as the carbon source. When gluconate and glycerol are used as the sole carbon source by Klebsiella oxytoca BW21, the ethanol yield is about 62 to 64%. Coutilization of both gluconate and glycerol in batch fermentation increased the yield of ethanol to about 78.7% and decreased by-product accumulation (such as acetate and 1,3-propanediol) substantially. Decreasing by-product formation by deleting the pta, frd, ldh, pflA, and pduC genes in strain BW21 increased the ethanol yield to 89.3% in the batch fermentation of a glycerol-gluconate mixture. These deletions produced the strain K. oxytoca WT26. However, the utilization rate of glycerol was significantly slower than that of gluconate in batch fermentation. In addition, substantial amounts of glycerol remain unutilized after gluconate was depleted in batch fermentation. Continuous fed-batch fermentation was used to solve the utilization rate mismatch problem for gluconate and glycerol. An ethanol yield of 97.2% was achieved in continuous fed-batch fermentation of these two substrates, and glycerol was completely used at the end of the fermentation. IMPORTANCE Gluconate is a biomass-derived degradation product, and glycerol can be obtained as a biodiesel byproduct. Compared to glucose, using them as the sole substrate is accompanied by the production of by-products. Our study shows that through pathway engineering and adoption of a fed-batch culture system, high-yield homoethanol production that usually can be achieved by using glucose as the substrate is achievable using gluconate and glycerol as cosubstrates. The same strategy is expected to be able to achieve homofermentative production of other products, such as lactate and 2,3-butanediol, which can be typically achieved using glucose as the substrate and inexpensive biodiesel-derived glycerol and biomass-derived gluconate as the cosubstrates.

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Fermentation of biodiesel-derived glycerol by Bacillus amyloliquefaciens: effects of co-substrates on 2,3-butanediol production

Cited by 49 publications

References 26 publications

Enhanced 2,3-butanediol production from biodiesel-derived glycerol by engineering of cofactor regeneration and manipulating carbon flux in Bacillus amyloliquefaciens

Enhanced 2,3-butanediol production from biodiesel-derived glycerol by engineering of cofactor regeneration and manipulating carbon flux in Bacillus amyloliquefaciens

Two-step production of gamma-aminobutyric acid from cassava powder using Corynebacterium glutamicum and Lactobacillus plantarum

Homoethanol Production from Glycerol and Gluconate Using Recombinant Klebsiella oxytoca Strains

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