Cloning and functional characterization of the UDP‐glucosyltransferase UgtB1 involved in sophorolipid production by <i>Candida bombicola</i> and creation of a glucolipid‐producing yeast strain

Saerens, Karen; Zhang, Jinxin; Saey, Lien; Bogaert, Inge Van; Soetaert, Wim

doi:10.1002/yea.1838

Cited by 73 publications

(83 citation statements)

References 56 publications

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“…All rights reserved 8 E n z y m e a s s a y s S. bombicola ATCC 22214 wild type, the ugtA1 deletion mutant A113 knocked out in the first glucosyltransferase (Saerens et al 2011a) and the ugtB1 deletion mutant B11 knocked out in the second glucosyltransferase (Saerens et al, 2011c) were grown overnight in 5mL 3C medium at 30°C and 200 rpm. The culture was used to inoculate 20 mL 3C medium and was incubated the same way for about 60 hours.…”

Section: Acc E P Ted P R E P R I Ntmentioning

confidence: 99%

Synthesis of bolaform biosurfactants by an engineered Starmerella bombicola yeast

Bogaert

Buyst

Martins

et al. 2016

Biotech & Bioengineering

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Bola-amphiphilic surfactants are molecules with fascinating properties. Their unique configuration consisting of a long hydrophobic spacer connecting two hydrophilic entities renders the molecule more water soluble than the average surfactant, but still allows formation of supramolecular structures. These properties make them extremely suitable for applications in in nanotechnology, electronics, and gene and drug delivery. In general, these compounds are obtained by chemical synthesis. We report here an efficient microbial production process for the fully green synthesis of bolaform surfactants. A sophorolipid-producing Starmerella bombicola yeast strain was disabled in its sophorolipid acetyltransferase and lactone esterase, which should logically result in synthesis of non-acetylated acidic sophorolipids; molecules with the classic amphiphilic structure. Surprisingly, also bolaform glycolipids were obtained, with an additional sophorose linked to the free carboxyl end of the acidic sophorolipids as confirmed by MS and NMR analysis. The obtained titers of 27.7 g/L total product are comparable to wild type values, and the novel molecules account for at least 74% of this. Bola-amphiphile biosynthesis proved to be attributed to the promiscuous activity of both UDP-glucosyltransferases UGTA1 and UGTB1 from the core sophorolipid pathway, displaying activity toward non-acetylated intermediates. The absence of acetyl groups seems to trigger formation of bolaform compounds starting from acidic sophorolipids. Hence, wild type S. bombicola produces these compounds only at marginal amounts in general not reaching detection limits. We created a strain knocked-out in its sophorolipid acetyltransferase and lactone esterase able to produce these novel compounds in economical relevant amounts, opening doors for the application of biological-derived bolaform structures. Biotechnol. Bioeng. 2016;113: 2644-2651. © 2016 Wiley Periodicals, Inc.

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Section: Acc E P Ted P R E P R I Ntmentioning

confidence: 99%

Synthesis of bolaform biosurfactants by an engineered Starmerella bombicola yeast

Bogaert

Buyst

Martins

et al. 2016

Biotech & Bioengineering

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“…The deletion fragments were transformed as previously reported by Saerens et al . (,b). The yeast cells (10 7 per ml) were inoculated into a 100‐ml Erlenmeyer flask containing 30 ml seed medium and incubated at 28–30°C, with shaking at 220 rpm for 2 days until the medium turned milky.…”

Section: Methodsmentioning

confidence: 99%

“…It has been reported that both glucosyltransferases were inhibited by their product SLs. Further, when the product concentration reached > 1 mM, the residual activity observed for UgtA1 was only 62% (Saerens et al ., ,b). Moreover, Haque et al .…”

Section: Introductionmentioning

confidence: 99%

Semicontinuous sophorolipid fermentation using a novel bioreactor with dual ventilation pipes and dual sieve‐plates coupled with a novel separation system

Zhang

Dan

Sun

et al. 2017

Microbial Biotechnology

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SummarySophorolipids (SLs) are biosurfactants with widespread applications. The yield and purity of SLs are two important factors to be considered during their commercial large‐scale production. Notably, SL accumulation causes an increase in viscosity, decrease in dissolved oxygen and product inhibition in the fermentation medium. This inhibits the further production and purification of SLs. This describes the development of a novel integrated system for SL production using Candida albicans O‐13‐1. Semicontinuous fermentation was performed using a novel bioreactor with dual ventilation pipes and dual sieve‐plates (DVDSB). SLs were separated and recovered using a newly designed two‐stage separation system. After SL recovery, the fermentation broth containing residual glucose and oleic acid was recycled back into the bioreactor. This novel approach considerably alleviated the problem of product inhibition and accelerated the rate of substrate utilization. Production of SLs achieved was 477 g l−1, while their productivity was 1.59 g l−1 h−1. Purity of SLs improved by 23.3%, from 60% to 74%, using DVDSB with the separation system. The conversion rate of carbon source increased from 0.5 g g−1 (in the batch fermentation) to 0.6 g g−1. These results indicated that the integrated system could improve the efficiency of production and purity of SLs.

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“…This activates the fatty acid for the addition of glucose by glucosyltransferase I, using uridine diphosphate (UDP)‐glucose as donor molecule (UgtA1; ). The second glucose is linked to the first via a glucosyl‐β‐(1,2)‐glucosyl linkage by glucosyltransferase II (UgtB1; ). The product of the second glycosylation, a nonacetylated SL, can be further decorated with one or two acetyl groups at the C6′ or C6″ position of the sophorose moiety.…”

Section: Sophorolipidsmentioning

confidence: 99%

“…Furthermore, the single knockout strategy resulted in a transformant capable of producing glucolipids rather than SLs; only the first of the two glucose units is attached to the fatty acid backbone. To this end, the ugtb1 gene was interrupted . Not only can the hydrophilic head be tailored through genetic engineering of the production host, also the hydrophobic tail of the SLs can be adjusted; by exchanging the C18‐prone Cyp52m1 responsible for hydroxylation of the fatty acid with C 16 ‐specific‐P450 enzymes, SLs with shorter fatty acid tails were obtained .…”

Section: Sophorolipidsmentioning

confidence: 99%

Yeast glycolipid biosurfactants

2017

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Edited by Wilhelm JustVarious yeasts, both conventional and exotic ones, are known to produce compounds useful to mankind. Ethanol is the most known of these compounds, but more complex molecules such as amphiphilic biosurfactants can also be derived from eukaryotic microorganisms at an industrially and commercially relevant scale. Among them, glycolipids are the most promising, due to their attractive properties and high product titers. Many of these compounds can be considered as secondary metabolites with a specific function for the host. Hence, a dedicated biosynthetic process enables regulation and combines pathways delivering the lipidic moiety and the hydrophilic carbohydrate part of the glycolipid. In this Review, we will discuss the biosynthetic and regulatory aspects of the yeast-derived sophorolipids, mannosylerythritol lipids, and cellobiose lipids, with special emphasis on the relation between glycolipid synthesis and the general lipid metabolism.

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Cloning and functional characterization of the UDP‐glucosyltransferase UgtB1 involved in sophorolipid production by Candida bombicola and creation of a glucolipid‐producing yeast strain

Cited by 73 publications

References 56 publications

Synthesis of bolaform biosurfactants by an engineered Starmerella bombicola yeast

Synthesis of bolaform biosurfactants by an engineered Starmerella bombicola yeast

Semicontinuous sophorolipid fermentation using a novel bioreactor with dual ventilation pipes and dual sieve‐plates coupled with a novel separation system

Yeast glycolipid biosurfactants

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