scholarly journals Optimization and Engineering of a Self-Sufficient CYP102 Enzyme from Bacillus amyloliquefaciens towards Synthesis of In-Chain Hydroxy Fatty Acids

Catalysts ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 665
Author(s):  
Li Zong ◽  
Yan Zhang ◽  
Zhengkang Shao ◽  
Yingwu Wang ◽  
Zheng Guo ◽  
...  

Cytochrome P450 (CYP) mediated enzymatic hydroxylation of fatty acids present a green alternative to chemical synthesis of hydroxy fatty acids (HFAs), which are high-value oleochemicals with various uses in materials industry and medical field. Although many CYPs require the presence of additional reductase proteins for catalytic activity, self-sufficient CYPs have their reductase partner naturally fused into their catalytic domain, leading to a greatly simplified biotransformation process. A recently discovered self-sufficient CYP, BAMF2522 from Bacillus amyloliquefaciens DSM 7, exhibits novel regioselectivity by hydroxylating in-chain positions of palmitic acid generating ω-1 to ω-7 HFAs, a rare regiodiversity profile among CYPs. Besides, F89I mutant of BAMF2522 expanded hydroxylation up to ω-9 position of palmitic acid. Here, we further characterize this enzyme by determining optimum temperature and pH as well as thermal stability. Moreover, using extensive site-directed and site-saturation mutagenesis, we obtained BAMF2522 variants that demonstrate greatly increased regioselectivity for in-chain positions (ω-4 to ω-9) of various medium to long chain fatty acids. Remarkably, when a six-residue mutant was reacted with palmitic acid, 84% of total product content was the sum of ω-7, ω-8 and ω-9 HFA products, the highest in-chain selectivity observed to date with a self-sufficient CYP. In short, our study demonstrates the potential of a recently identified CYP and its mutants for green and sustainable production of a variety of in-chain hydroxy enriched HFAs.

1969 ◽  
Vol 22 (4) ◽  
pp. 1015 ◽  
Author(s):  
TJ Heath ◽  
L N Hill

Normal sheep and sheep with fistulae of the bile duct or the thoracic duct were used to provide quantitative information on the movement of fatty acids into and out of the intestine. The operation used to gain access to the thoracic duct did not cause any significant alteration in the absorption of either [14C]tripalmitin injected into the rumen or [14C]palmitic acid injected into the duodenum. Normal sheep absorbed the major fatty acids oleic (92'1�1'3%), palmitic (87�3�5�0%), and stearic acids (93' 3� 1� 4%) with almost equal efficiency, and the absorption of labelled tri-palmitin injected into the rumen did not alter as the intake of fatty acids increased from 12g/day (90�1�2�3%) to 44g/day (90�1�1�3%).


1964 ◽  
Vol 206 (1) ◽  
pp. 153-158 ◽  
Author(s):  
Martin Gold ◽  
John J. Spitzer

The myocardial and renal removal and oxidation of plasma free fatty acids (FFA) were studied in dogs receiving an infusion of I-C14-palmitate, oleate, linoleate, or octanoate. The myocardium extracted about 40% of the total arterial FFA, and some was oxidized to CO2. Blood C14O2 was consistently higher in the coronary sinus than in the artery. The specific activity of the coronary sinus C14O2 was higher than that of the arterial C14O2 in dogs infused with long-chain fatty acids. The specific activity of arterial C14O2 was higher than that of the coronary sinus C14O2 when octanoate was infused. There were no differences observed in FFA extraction and C14O2 production among the different groups of animals receiving the various long-chain fatty acids. In gas chromatographic studies palmitic acid was the only FFA with a consistent net removal by the kidney. Approximately 9% of the arterial radiopalmitate was removed by this organ in dogs receiving infusions of palmitic acid-1-C14. Simultaneously, C14O2 was produced by the kidney. A consistent removal of oleate-1-C14 or a significant production of C14O2 was not found. Infusion of octanoate-1-C14 led to a marked release of C14O2 by the kidney, with greatly increased CO2 specific activity in the renal vein.


1977 ◽  
Vol 168 (2) ◽  
pp. 261-269 ◽  
Author(s):  
Paul Bolton ◽  
John L. Harwood

1. Fatty acid synthesis was studied in microsomal preparations from germinating pea (Pisum sativum). 2. The preparations synthesized a mixture of saturated fatty acids up to a chain length of C24 from [14C]malonyl-CoA. 3. Whereas hexadecanoic acid was made de novo, octadecanoic acid and icosanoic acid were synthesized by elongation. 4. The products formed during [14C]malonyl-CoA incubation were analysed, and unesterified fatty acids and polar lipids were found to be major products. [14C]Palmitic acid represented a high percentage of the acyl-carrier protein esters, whereas 14C-labelled very-long-chain fatty acids were mainly present as unesterified fatty acids. CoA esters were minor products. 5. The addition of exogenous lipids to the incubation system usually resulted in stimulation of [14C]malonyl-CoA incorporation into fatty acids. The greatest stimulation was obtained with dipalmitoyl phosphatidylcholine. Both exogenous palmitic acid and dipalmitoyl phosphatidylcholine increased the amount of [14C]-stearic acid synthesized, relative to [14C]palmitic acid. Addition of stearic acid increased the amount of [14C]icosanoic acid formed. 6. [14C]Stearic acid was elongated more effectively to icosanoic acid than [14C]stearoyl-CoA, and its conversion was not decreased by addition of unlabelled stearoyl-CoA. 7. Incorporation of [14C]malonyl-CoA into fatty acids was markedly decreased by iodoacetamide and 5,5′-dithiobis-(2-nitrobenzoic acid). Palmitate elongation was sensitive to arsenite addition, and stearate elongation to the presence of Triton X-100 or fluoride. The action of fluoride was not, apparently, due to chelation. 8. The microsomal preparations differed from soluble fractions from germinating pea in (a) synthesizing very-long-chain fatty acids, (b) not utilizing exogenous palmitate–acyl-carrier protein as a substrate for palmitate elongation and (c) having fatty acid synthesis stimulated by the addition of certain complex lipids.


2011 ◽  
Vol 58 (1) ◽  
Author(s):  
Adam Choma ◽  
Iwona Komaniecka

Lipopolysaccharides of seven Bradyrhizobium strains and three whole-cell fatty acid preparations from bacteria isolated from nodules of Sarothamnus scoparius (common broom) were studied for the presence of very long chain (ω-1)-hydroxy fatty acids. Several such fatty acids were identified. Among them, straight-chain as well as mono- and dimethyl branched acids with chains in the range from 26 to 34 carbon atoms were found. Pyrrolidides and 4,4-dimethyloxazoline derivatives were used to determine the branching position. Carbons at the (ω-10) and/or (ω-11) positions in alkyl chains were points of attachment of methyl groups. These data complete the structure of bradyrhizobial lipid A with important details. The obtained results can be applied in the chemotaxonomy of Bradyrhizobium.


1976 ◽  
Vol 3 (6) ◽  
pp. 635-636
Author(s):  
R J Stretton ◽  
R K Dart

A number of strains purporting to belong to the species Sporothrix schenckii were examined for their fatty acid content. The majority of the strains were isolated from cases of sporotrichosis. Two strains were reputedly saprophytic. In all cases except the two saprophytic ones the major fatty acid was a C18 diene. Considerable amounts of palmitic acid and C18 monoene were found in all strains.


1996 ◽  
Vol 76 (03) ◽  
pp. 369-371 ◽  
Author(s):  
T A B Sanders ◽  
G J Miller ◽  
Tamara de Grassi ◽  
Najat Yahia

SummaryFactor VII coagulant activity (FVIIc) is associated with an increased risk of fatal ischaemic heart disease (IHD). Several reports have suggested that dietary fat intake or hypertriglyceridaemia are associated with elevated levels of FVII. This study demonstrates that an intake of long-chain fatty acids sufficient to induce postprandial lipaemia in healthy subjects leads to a substantial elevation in both FVIIc and the concentration of FVII circulating in the activated form. Such an increase in FVIIc could not be induced by medium-chain triglycerides. These results suggest that the consumption of a sufficient amount of long-chain triglycerides to induce postprandial lipaemia induces the activation of FVII.


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