scholarly journals Genetic engineering of the precursor supply pathway for the overproduction of the nC14-surfactin isoform with promising MEOR applications

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Fangxiang Hu ◽  
Weijie Cai ◽  
Junzhang Lin ◽  
Weidong Wang ◽  
Shuang Li
Keyword(s):  
Bacillus Subtilis ◽  
Fatty Acid ◽  
Surface Activity ◽  
Acyl Carrier Protein ◽  
Emulsification Index ◽  
Fatty Acid Chain ◽  
Fatty Acid Precursor ◽  
Surfactin Production ◽  
Acid Precursor ◽  
Precursor Supply

Abstract Background Surfactin, a representative biosurfactant of lipopeptide mainly produced by Bacillus subtilis, consists of a cyclic heptapeptide linked to a β-hydroxy fatty acid chain. The functional activity of surfactin is closely related to the length and isomerism of the fatty acid chain. Results In this study, the fatty acid precursor supply pathway in Bacillus subtilis 168 for surfactin production was strengthened through two steps. Firstly, pathways competing for the precursors were eliminated with inactivation of pps and pks. Secondly, the plant medium-chain acyl-carrier protein (ACP) thioesterase (BTE) from Umbellularia californica was overexpressed. As a result, the surfactin titer after 24 h of cultivation improved by 34%, and the production rate increased from 0.112 to 0.177 g/L/h. The isoforms identified by RP-HPLC and GC–MS showed that the proportion of nC14-surfactin increased 6.4 times compared to the control strain. A comparison of further properties revealed that the product with more nC14-surfactin had higher surface activity and better performance in oil-washing. Finally, the product with more nC14-surfactin isoform had a higher hydrocarbon-emulsification index, and it increased the water-wettability of the oil-saturated silicate surface. Conclusion The obtained results identified that enhancing the supply of fatty acid precursor is very essential for the synthesis of surfactin. At the same time, this study also proved that thioesterase BTE can promote the production of nC14-surfactin and experimentally demonstrated its higher surface activity and better performance in oil-washing. These results are of great significance for the MEOR application of surfactin. Graphic abstract

scholarly journals Overexpression of the Plant Medium-chain Acyl-carrier Protein Thioesterase BTE for the Overproduction of the nC14-surfactin Isoform with Promising MEOR Applications

2021 ◽  
Author(s):  
fangxiang hu ◽  
Weijie Cai ◽  
Junzhang Lin ◽  
Weidong Wang ◽  
Shuang Li
Keyword(s):  
Fatty Acid ◽  
Hydroxy Fatty Acid ◽  
Acyl Carrier Protein ◽  
Carrier Protein ◽  
Medium Chain ◽  
Emulsification Index ◽  
Fatty Acid Chain ◽  
Umbellularia Californica ◽  
Chain Acyl ◽  
Surfactin Production

Abstract BackgroundSurfactin, a representative biosurfactant of popeptide mainly produced by Bacillus subtilis, consists of a cyclic heptapeptide linked to a β-hydroxy fatty acid chain. The functional activity of surfactin is closely related to the length and isomerism of the fatty acid chain. ResultsIn this study, the plant medium-chain acyl-carrier protein (ACP) thioesterase (BTE) from Umbellularia californica was overexpressed in a recombinant surfactin production strain based on B. subtilis 168. As a result, the surfactin yield after 24 h of cultivation improved by 23%, and the production rate increased from 0.112 to 0.177 g/L/h. The isoforms identified by RP-HPLC and GC-MS showed that the proportion of nC14-surfactin increased 6.4 times compared to the control strain. A comparison of further properties revealed that the product with more nC14-surfactin had higher surface activity and better performance in oil-washing. Finally, the product with more nC14-surfactin isoform had a higher hydrocarbon-emulsification index, and it increased the water-wettability of the oil-saturated silicate surface. ConclusionThe obtained results provide an original approach to modify the fatty acid chain of surfactin and further demonstrate the importance of the length and isomerism of the β-hydroxy fatty acid chain for the MEOR application of surfactin.

Biosynthesis of the Furanoacetylene Phytoalexin Wyerone in Vicia faba

1986 ◽  
Vol 41 (1-2) ◽  
pp. 34-38 ◽  
Author(s):  
Nidhal A. Al-Douri ◽  
Paul M. Dewick
Keyword(s):  
Fatty Acid ◽  
Vicia Faba ◽  
Sodium Acetate ◽  
Biosynthetic Pathway ◽  
Broad Bean ◽  
Feeding Experiments ◽  
Fatty Acid Precursor ◽  
Acid Precursor

Abstract Feeding experiments using 13C-labelled sodium acetate precursors in CuCl2-treated broad bean (Vicia faba) cotyledons have demonstrated that the furanoacetylene phytoalexin wyerone is biosynthetically derived from seven intact acetate units. A further experiment using sodium [2H3]acetate indicated the head of the chain, and showed the chain is analogous to that of a fatty acid precursor, any chain shortening process from postulated C18 precursors occurring from the carboxyl end. Incorporations of oleate and linoleate were, however, regarded as insufficient to prove the involvement of these compounds in the biosynthetic pathway.

scholarly journals Effects of Different Cultivation Parameters on the Production of Surfactin Variants by a Bacillus subtilis Strain

Molecules ◽  
2018 ◽  
Vol 23 (10) ◽  
pp. 2675 ◽  
Author(s):  
Attila Bartal ◽  
Aruna Vigneshwari ◽  
Bettina Bóka ◽  
Mónika Vörös ◽  
István Takács ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Metal Ions ◽  
Biological Activities ◽  
Wide Spectrum ◽  
Carbon Sources ◽  
Bacillus Species ◽  
Subtilis Strain ◽  
Fatty Acid Chain ◽  
Characteristic Features ◽  
Surfactin Production

Surfactins are lipopeptide-type biosurfactants produced mainly by Bacillus species, consisting of a peptide loop of seven amino acids and a hydrophobic fatty acid chain (C12–C16). These molecules have been proven to exhibit various biological activities; thus, their therapeutic and environmental applications are considered. Within the surfactin lipopeptide family, there is a wide spectrum of different homologues and isomers; to date, more than 30 variants have been described. Since the newest members of these lipopeptides were described recently, there is no information that is available on their characteristic features, e.g., the dependence of their production from different cultivation parameters. This study examined the effects of both the different carbon sources and various metal ions on the surfactin production of a selected B. subtilis strain. Among the applied carbon sources, fructose and xylose had the highest impacts on the ratio of the different variants, regarding both the peptide sequences and the lengths of the fatty acids. Furthermore, the application of metal ions Mn2+, Cu2+ and Ni2+ in the media completely changed the surfactin variant compositions of the fermenting broths leading to the appearance of methyl esterified surfactin forms, and resulted in the appearance of novel surfactin variants with fatty acid chains containing no more than 11 carbon atoms.

scholarly journals Exogenous Isoleucine and Fatty Acid Shortening Ensure the High Content of Anteiso-C15:0 Fatty Acid Required for Low-Temperature Growth of Listeria monocytogenes

2005 ◽  
Vol 71 (12) ◽  
pp. 8002-8007 ◽  
Author(s):  
Kun Zhu ◽  
Xiang Ding ◽  
Mudcharee Julotok ◽  
Brian J. Wilkinson
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Listeria Monocytogenes ◽  
Low Temperature ◽  
Fatty Acid Profile ◽  
Acyl Carrier Protein ◽  
Critical Role ◽  
Chain Fatty Acid ◽  
Fatty Acid Chain ◽  
Branched Chain

ABSTRACT Previous studies have demonstrated that the branched-chain fatty acid anteiso-C15:0 plays a critical role in the growth of Listeria monocytogenes at low temperatures by ensuring sufficient membrane fluidity. Studies utilizing a chemically defined minimal medium revealed that the anteiso fatty acid precursor isoleucine largely determined the fatty acid profile and fatty acid response of the organism to lowered growth temperature. When isoleucine was sufficient, the fatty acid profile was very uniform, with anteiso fatty acids comprising up to 95% of total fatty acid, and the major fatty acid adjustment to low temperature was fatty acid chain shortening, which resulted in an increase of anteiso-C15:0 solely at the expense of anteiso-C17:0. When isoleucine was not supplied, the fatty acid profile became more complex and was readily modified by leucine, which resulted in a significant increase of corresponding iso fatty acids and an inability to grow at 10°C. Under this condition, the increase of anteiso-C15:0 at low temperature resulted from the combined effect of increasing the anteiso:iso ratio and chain shortening. A branched-chain α-keto acid dehydrogenase-defective strain largely lost the ability to increase the anteiso:iso ratio. Cerulenin, an inhibitor of β-ketoacyl-acyl carrier protein synthase (FabF), induced a similar fatty acid chain shortening as low temperature did. We propose that the anteiso precursor preferences of enzymes in the branched-chain fatty acid biosynthesis pathway ensure a high production of anteiso fatty acids, and cold-regulated chain shortening results in a further increase of anteiso-C15:0 at the expense of anteiso-C17:0.

Selectivity in a barren landscape: the P450BioI–ACP complex

2010 ◽  
Vol 38 (4) ◽  
pp. 934-939 ◽  
Author(s):  
Max J. Cryle
Keyword(s):  
Fatty Acid ◽  
Protein Complex ◽  
Acyl Carrier Protein ◽  
Protein Complexes ◽  
Cytochromes P450 ◽  
Carrier Protein ◽  
Pimelic Acid ◽  
Fatty Acid Chain ◽  
Fatty Acid Hydroxylation ◽  
Oxidative Transformations

The cytochromes P450 (P450s) are a superfamily of oxidative haemoproteins that are capable of catalysing a vast range of oxidative transformations, including the oxidation of unactivated alkanes, often with high stereo- and regio-selectivity. Fatty acid hydroxylation by P450s is widespread across both bacteria and higher organisms, with the sites of oxidation and specificity of oxidation varying from system to system. Several key examples are discussed in the present article, with the focus on P450BioI (CYP107H1), a biosynthetic P450 found in the biotin operon of Bacillus subtilis. The biosynthetic function of P450BioI is the formation of pimelic acid, a biotin precursor, via a multiple-step oxidative cleavage of long-chain fatty acids. P450BioI is a member of an important subgroup of P450s that accept their substrates not free in solution, but rather presented by a separate carrier protein. Structural characterization of the P450BioI–ACP (acyl-carrier protein) complex has recently been performed, which has revealed the basis for the oxidation of the centre of the fatty acid chain. The P450BioI–ACP structure is the first such P450–carrier protein complex to be characterized structurally, with important implications for other biosynthetically intriguing P450–carrier protein complexes.

scholarly journals Transcriptional and functional characterization of the gene encoding acyl carrier protein in Bacillus subtilis

Microbiology ◽  
2010 ◽  
Vol 156 (2) ◽  
pp. 484-495 ◽  
Author(s):  
Mariano A. Martinez ◽  
Diego de Mendoza ◽  
Gustavo E. Schujman
Keyword(s):  
Bacillus Subtilis ◽  
Fatty Acid ◽  
Molecular Mechanisms ◽  
Fatty Acid Biosynthesis ◽  
Acyl Carrier Protein ◽  
Functional Characterization ◽  
Carrier Protein ◽  
Normal Fatty Acid ◽  
Gram Positive

Acyl carrier protein (ACP) is a universal and highly conserved carrier of acyl intermediates during fatty acid biosynthesis. The molecular mechanisms of regulation of the acpP structural gene, as well as the function of its gene product, are poorly characterized in Bacillus subtilis and other Gram-positive organisms. Here, we report that transcription of acpP takes place from two different promoters: PfapR and PacpP. Expression of acpP from PfapR is coordinated with a cluster of genes involved in lipid synthesis (the fapR operon); the operon consists of fapR-plsX-fabD-fabG-acpP. PacpP is located immediately upstream of the acpP coding sequence, and is necessary and sufficient for normal fatty acid synthesis. We also report that acpP is essential for growth and differentiation, and that ACP localizes in the mother-cell compartment of the sporangium during spore formation. These results provide the first detailed characterization of the expression of the ACP-encoding gene in a Gram-positive bacterium, and highlight the importance of this protein in B. subtilis physiology.

scholarly journals Response of Bacillus subtilis to Cerulenin and Acquisition of Resistance

2001 ◽  
Vol 183 (10) ◽  
pp. 3032-3040 ◽  
Author(s):  
Gustavo E. Schujman ◽  
Keum-Hwa Choi ◽  
Silvia Altabe ◽  
Charles O. Rock ◽  
Diego de Mendoza
Keyword(s):  
Bacillus Subtilis ◽  
Fatty Acid ◽  
Reporter Gene ◽  
Fatty Acid Synthesis ◽  
Acyl Carrier Protein ◽  
Acyl Chain ◽  
Acid Synthesis ◽  
Covalent Modification ◽  
Chain Elongation ◽  
Growth Defect

ABSTRACT Cerulenin is a fungal mycotoxin that potently inhibits fatty acid synthesis by covalent modification of the active site thiol of the chain-elongation subtypes of β-ketoacyl-acyl carrier protein (ACP) synthases. The Bacillus subtilis fabF (yjaY) gene (fabFb ) encodes an enzyme that catalyzes the condensation of malonyl-ACP with acyl-ACP to extend the growing acyl chain by two carbons. There were two mechanisms by which B. subtilis adapted to exposure to this antibiotic. First, reporter gene analysis demonstrated that transcription of the operon containing the fabF gene increased eightfold in response to a cerulenin challenge. This response was selective for the inhibition of fatty acid synthesis, since triclosan, an inhibitor of enoyl-ACP reductase, triggered an increase in fabF reporter gene expression while nalidixic acid did not. Second, spontaneous mutants arose that exhibited a 10-fold increase in the MIC of cerulenin. The mutation mapped at the B. subtilis fabF locus, and sequence analysis of the mutant fabF allele showed that a single base change resulted in the synthesis of FabFb[I108F]. The purified FabFb and FabFb[I108F] proteins had similar specific activities with myristoyl-ACP as the substrate. FabFb exhibited a 50% inhibitory concentration (IC50) of cerulenin of 0.1 μM, whereas the IC50 for FabFb[I108] was 50-fold higher (5 μM). These biochemical data explain the absence of an overt growth defect coupled with the cerulenin resistance phenotype of the mutant strain.

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