Whole-cell based solvent-free system for one-pot production of biodiesel from waste grease

Li, Aitao; Ngo, Thao P.N.; Yan, Jinyong; Tian, Kaiyuan; Li, Zhi

doi:10.1016/j.biortech.2012.03.034

Cited by 49 publications

(29 citation statements)

References 14 publications

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“…1). Previous WCB reports highlighted similar observations and recommended cell permeation to cope with mass transfer issues across cell membranes [6], [11], [24], [27], [36].…”

Section: Resultssupporting

confidence: 56%

Short-Chain Flavor Ester Synthesis in Organic Media by an E. coli Whole-Cell Biocatalyst Expressing a Newly Characterized Heterologous Lipase

et al. 2014

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Short-chain aliphatic esters are small volatile molecules that produce fruity and pleasant aromas and flavors. Most of these esters are artificially produced or extracted from natural sources at high cost. It is, however, possible to ‘naturally’ produce these molecules using biocatalysts such as lipases and esterases. A gene coding for a newly uncovered lipase was isolated from a previous metagenomic study and cloned into E. coli BL21 (DE3) for overexpression using the pET16b plasmid. Using this recombinant strain as a whole-cell biocatalyst, short chain esters were efficiently synthesized by transesterification and esterification reactions in organic media. The recombinant lipase (LipIAF5-2) showed good affinity toward glyceryl trioctanoate and the highest conversion yields were obtained for the transesterification of glyceryl triacetate with methanol. Using a simple cetyl-trimethylammonium bromide pretreatment increased the synthetic activity by a six-fold factor and the whole-cell biocatalyst showed the highest activity at 40°C with a relatively high water content of 10% (w/w). The whole-cell biocatalyst showed excellent tolerance to alcohol and short-chain fatty acid denaturation. Substrate affinity was equally effective with all primary alcohols tested as acyl acceptors, with a slight preference for methanol. The best transesterification conversion of 50 mmol glyceryl triacetate into isoamyl acetate (banana fragrance) provided near 100% yield after 24 hours using 10% biocatalyst loading (w/w) in a fluidized bed reactor, allowing recycling of the biocatalyst up to five times. These results show promising potential for an industrial approach aimed at the biosynthesis of short-chain esters, namely for natural flavor and fragrance production in micro-aqueous media.

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“…1). Previous WCB reports highlighted similar observations and recommended cell permeation to cope with mass transfer issues across cell membranes [6], [11], [24], [27], [36].…”

Section: Resultssupporting

confidence: 56%

Short-Chain Flavor Ester Synthesis in Organic Media by an E. coli Whole-Cell Biocatalyst Expressing a Newly Characterized Heterologous Lipase

et al. 2014

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“…The highest biodiesel yield was derived using a ratio of 6:1 (methanol:oil), demonstrating better methanol tolerance in this integrated system than in other previous biocatalytic systems (generally 3:1 to 4:1) based on immobilized enzymes or whole cell catalysts [22,23]. Further increase of the amount of methanol (9:1) did not improve biodiesel production, likely because of the significant loss of lipase activity.…”

Section: Resultsmentioning

confidence: 75%

“…Nonetheless, it was estimated that this integrated biodiesel production system contained 50% to 100% water relative to oil content. In previous immobilized enzyme- or whole cell lipase-based biodiesel production systems [22,23], water content was generally limited to very low level (<3%) to achieve satisfactory biodiesel yield through the addition of anhydrous molecular sieves or silica gels as water absorbent agents. The significantly enhanced water tolerance of this integrated system could significantly simplify the operation process and further reduce costs.…”

Section: Resultsmentioning

confidence: 99%

Integrated lipase production and in situ biodiesel synthesis in a recombinant Pichia pastoris yeast: an efficient dual biocatalytic system composed of cell free enzymes and whole cell catalysts

Yan

Zheng

et al. 2014

Biotechnol Biofuels

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BackgroundLipase-catalyzed biotransformation of acylglycerides or fatty acids into biodiesel via immobilized enzymes or whole cell catalysts has been considered as one of the most promising methods to produce renewable and environmentally friendly alternative liquid fuels, thus being extensively studied so far. In all previously pursued approaches, however, lipase enzymes are prepared in an independent process separated from enzymatic biodiesel production, which would unavoidably increase the cost and energy consumption during industrial manufacture of this cost-sensitive energy product. Therefore, there is an urgent need to develop novel cost-effective biocatalysts and biocatalytic processes with genuine industrial feasibility.ResultInspired by the consolidated bioprocessing of lignocellulose to generate bioethanol, an integrated process with coupled lipase production and in situ biodiesel synthesis in a recombinant P. pastoris yeast was developed in this study. The novel and efficient dual biocatalytic system based on Thermomyces lanuginosus lipase took advantage of both cell free enzymes and whole cell catalysts. The extracellular and intracellular lipases of growing yeast cells were simultaneously utilized to produce biodiesel from waste cooking oils in situ and in one pot. This integrated system effectively achieved 58% and 72% biodiesel yield via concurrent esterified-transesterified methanolysis and stepwise hydrolysis-esterification at 3:1 molar ratio between methanol and waste cooking oils, respectively. Further increasing the molar ratio of methanol to waste cooking oils to 6:1 led to an 87% biodiesel yield using the stepwise strategy. Both water tolerance and methanol tolerance of this novel system were found to be significantly improved compared to previous non-integrated biodiesel production processes using separately prepared immobilized enzymes or whole cell catalysts.ConclusionWe have proposed a new concept of integrated biodiesel production. This integrated system couples lipase production to lipase-catalyzed biodiesel synthesis in one pot. The proof-of-concept was established through construction of a recombinant P. pastoris yeast strain that was able to grow, overexpress T. lanuginosus lipase, and efficiently catalyze biodiesel production from fed waste cooking oils and methanol simultaneously. This simplified single-step process represents a significant advance toward achieving economical production of biodiesel at industrial scale via a ‘green’ biocatalytic route.

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“…FFAs can be produced by introducing an acyl-acyl carrier protein (ACP) thioesterase (TE) gene into E. coli. The presence of the acyl-ACP TE breaks the fatty acid elongation cycle and releases FFAs (Lu et al 2008;Li et al 2012a;Zhang et al 2011;Youngquist et al 2013). Ingram and colleagues (1977) used propionate to induce synthesis of odd-chain-length fatty acids by wild type Escherichia coli K-12.…”

Section: Introductionmentioning

confidence: 98%

Engineering Escherichia coli for odd straight medium chain free fatty acid production

San

2014

Appl Microbiol Biotechnol

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Microbial biosynthesis of free fatty acids (FFAs) can be achieved by introducing an acyl-acyl carrier protein thioesterase gene into Escherichia coli. The engineered E. coli usually produced even chain FFAs. In this study, propionyl-CoA synthetase (prpE) from Salmonella enterica was overexpressed in two efficient even chain FFAs producers, ML103 (pXZM12) carrying the acyl-ACP thioesterase gene from Umbellularia californica and ML103 (pXZ18) carrying the acyl-ACP thioesterase gene from Ricinus communis combined with supplement of extracellular propionate. With these metabolically engineered E. coli, the odd straight chain FFAs, undecanoic acid (C11:0), tridecanoic acid (C13:0), and pentadecanoic acid (C15:0) were produced from glucose and propionate. The highest total odd straight chain FFAs produced by ML103 (pXZM12, pBAD-prpE) reached 276 mg/l with a ratio of 23.43 % of the total FFAs. In ML103 (pXZ18, pBAD-prpE), the highest total odd straight chain FFAs accumulated to 297 mg/l, and the ratio reached 17.68 % of the total FFAs. Due to the different substrate specificity of the acyl-ACP thioesterases, the major odd straight chain FFA components of ML103 (pXZM12, pBAD-prpE) were undecanoic acid and tridecanoic acid, while the ML103 (pXZ18, pBAD-prpE) preferred pentadecanoic acid.

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Whole-cell based solvent-free system for one-pot production of biodiesel from waste grease

Cited by 49 publications

References 14 publications

Short-Chain Flavor Ester Synthesis in Organic Media by an E. coli Whole-Cell Biocatalyst Expressing a Newly Characterized Heterologous Lipase

Short-Chain Flavor Ester Synthesis in Organic Media by an E. coli Whole-Cell Biocatalyst Expressing a Newly Characterized Heterologous Lipase

Integrated lipase production and in situ biodiesel synthesis in a recombinant Pichia pastoris yeast: an efficient dual biocatalytic system composed of cell free enzymes and whole cell catalysts

Engineering Escherichia coli for odd straight medium chain free fatty acid production

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