Genome-wide identification of membrane-bound fatty acid desaturase genes in Gossypium hirsutum and their expressions during abiotic stress

The consensus sequence of the third histidine box of a range of Δ5, Δ6, Δ8 and sphingolipid desaturases differs from that of the membrane-bound non-fusion Δ12 and Δ15 desaturases in the presence of glutamine instead of histidine. We have used site-directed mutagenesis to determine the importance of glutamine and other residues of the third histidine box and created a chimaeric enzyme to determine the ability of the Cyt b5 fusion domain from the plant sphingolipid desaturase to substitute for the endogenous domain of the Δ6 desaturase.

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Modulation of GmFAD3 expression alters responses to abiotic stress in soybean

10.22541/au.160569953.30220980/v1 ◽

2020 ◽

Author(s):

Ajay Singh ◽

Mahesh Kumar ◽

Susheel Raina ◽

Milind Ratnaparkhe ◽

Jagadish Rane ◽

...

Keyword(s):

Heat Stress ◽

Abiotic Stress ◽

Fatty Acid ◽

Stress Tolerance ◽

Salinity Stress ◽

Stress Responses ◽

Fatty Acid Desaturase ◽

Stress Conditions ◽

Omega 3 ◽

Soybean Plants

FAD3 play important roles in modulating membrane fluidity in response to various abiotic stresses. However, a comprehensive analysis of FAD3 in drought, salinity and heat stress tolerance is lacking in soybean. The present study assessed the functional role of fatty acid desaturase 3 to abiotic stress responses in soybean. We used Bean Pod Mottle Virus -based vector to alter expression of Glycine max omega-3 fatty acid desaturase . Higher levels of recombinant BPMV-GmFAD3 transcripts were detected in overexpressing soybean plants. Overexpression of GmFAD3 in soybean resulted in increased levels of jasmonic acid and higher expression of GmWRKY54 as compared to mock-inoculated, vector-infected and FAD3-silenced soybean plants under drought and salinity stress conditions. FAD3 overexpressing plants showed higher levels of chlorophyll content, leaf SPAD value, relative water content, chlorophyll fluorescence, transpiration rate, carbon assimilation rate, proline content and also cooler canopy under drought and salinity stress conditions as compared to mock-inoculated, vector-infected and FAD3-silenced soybean plants. Results from current study revealed that GmFAD3 overexpressing soybean plants exhibited drought and salinity stress tolerance although tolerance to heat stress was reduced. On the other hand, soybean plants silenced for GmFAD3 exhibited tolerance to heat stress, but were vulnerable to drought and salinity stress

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Genome-wide identification and expression profiling of the fatty acid desaturase gene family in the silkworm, Bombyx mori

Genetics and Molecular Research ◽

10.4238/2014.may.13.2 ◽

2014 ◽

Vol 13 (2) ◽

pp. 3747-3760 ◽

Cited By ~ 7

Author(s):

Q.M. Chen ◽

D.J. Cheng ◽

S.P. Liu ◽

Z.G. Ma ◽

X. Tan ◽

...

Keyword(s):

Fatty Acid ◽

Gene Family ◽

Bombyx Mori ◽

Expression Profiling ◽

Fatty Acid Desaturase ◽

Desaturase Gene ◽

Genome Wide ◽

Silkworm Bombyx Mori ◽

Fatty Acid Desaturase Gene

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A novel Delta12-fatty acid desaturase gene from methylotrophic yeast Pichia pastoris GS115.

Acta Biochimica Polonica ◽

10.18388/abp.2006_3303 ◽

2006 ◽

Vol 53 (4) ◽

pp. 753-759 ◽

Cited By ~ 15

Author(s):

D Sh Wei ◽

M Ch Li ◽

X X Zhang ◽

H Zhou ◽

L J Xing

Keyword(s):

Fatty Acid ◽

Functional Characterization ◽

Fatty Acid Desaturase ◽

Methylotrophic Yeast ◽

Pastoris Gs115 ◽

Desaturase Gene ◽

Methylotrophic Yeast Pichia Pastoris ◽

Membrane Bound ◽

Fatty Acid Desaturase Gene

The methylotrophic yeast Pichia pastoris GS115, a widely used strain in production of various heterologous proteins, especially membrane-bound enzymes, can also produce linoleic and linolenic acids, which indicates the existence of membrane-bound Delta12 and Delta15-fatty acid desaturases. This paper describes the cloning and functional characterization of a novel Delta12-fatty acid desaturase gene from this methylotrophic yeast. The open reading frame of the gene (named Pp-FAD12) is 1263 bp in size and encodes a 420-amino-acid peptide. The deduced Pp-FAD12 protein shows high identity (50-67%) with Delta12-fatty acid desaturases from other fungi. It also shows a high identity (57%) with Delta15-fatty acid desaturase (named Sk-FAD15) from Saccharomyces kluyveri. Expression of Pp-FAD12 in polyunsaturated fatty acids non-producing yeast Saccharomyces cerevisiae demonstrated that its product converted oleic acid (18 : 1) to linoleic acid (18 : 2). This result suggests that Pp-FAD12 encodes a novel Delta12-fatty acid desaturase in P. pastoris GS115. This is the first report about the cloning and functional characterization of Delta12-fatty acid desaturase gene in methylotrophic yeast.

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Homology modeling and docking studies of a Δ9-fatty acid desaturase from a Cold-tolerantPseudomonassp. AMS8

PeerJ ◽

10.7717/peerj.4347 ◽

2018 ◽

Vol 6 ◽

pp. e4347 ◽

Cited By ~ 2

Author(s):

Lawal Garba ◽

Mohamad Ariff Mohamad Yussoff ◽

Khairul Bariyyah Abd Halim ◽

Siti Nor Hasmah Ishak ◽

Mohd Shukuri Mohamad Ali ◽

...

Keyword(s):

Fatty Acid ◽

Homology Modeling ◽

Palmitic Acid ◽

Fatty Acid Desaturase ◽

Three Dimensional ◽

Docking Studies ◽

Fatty Acid Desaturases ◽

Protein Model ◽

Cold Tolerant ◽

Membrane Bound

Membrane-bound fatty acid desaturases perform oxygenated desaturation reactions to insert double bonds within fatty acyl chains in regioselective and stereoselective manners. The Δ9-fatty acid desaturase strictly creates the first double bond between C9 and 10 positions of most saturated substrates. As the three-dimensional structures of the bacterial membrane fatty acid desaturases are not available, relevant information about the enzymes are derived from their amino acid sequences, site-directed mutagenesis and domain swapping in similar membrane-bound desaturases. The cold-tolerantPseudomonassp. AMS8 was found to produce high amount of monounsaturated fatty acids at low temperature. Subsequently, an active Δ9-fatty acid desaturase was isolated and functionally expressed inEscherichia coli. In this paper we report homology modeling and docking studies of a Δ9-fatty acid desaturase from a Cold-tolerantPseudomonassp. AMS8 for the first time to the best of our knowledge. Three dimensional structure of the enzyme was built using MODELLER version 9.18 using a suitable template. The protein model contained the three conserved-histidine residues typical for all membrane-bound desaturase catalytic activity. The structure was subjected to energy minimization and checked for correctness using Ramachandran plots and ERRAT, which showed a good quality model of 91.6 and 65.0%, respectively. The protein model was used to preform MD simulation and docking of palmitic acid using CHARMM36 force field in GROMACS Version 5 and Autodock tool Version 4.2, respectively. The docking simulation with the lowest binding energy, −6.8 kcal/mol had a number of residues in close contact with the docked palmitic acid namely, Ile26, Tyr95, Val179, Gly180, Pro64, Glu203, His34, His206, His71, Arg182, Thr85, Lys98 and His177. Interestingly, among the binding residues are His34, His71 and His206 from the first, second, and third conserved histidine motif, respectively, which constitute the active site of the enzyme. The results obtained are in compliance with thein vivoactivity of the Δ9-fatty acid desaturase on the membrane phospholipids.

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Modulation of GmFAD3 Expression Alters Abiotic Stress Responses in Soybean

10.21203/rs.3.rs-1212971/v1 ◽

2022 ◽

Author(s):

Ajay K Singh ◽

Susheel Kumar Raina ◽

Mahesh Kumar ◽

Lalitkumar Aher ◽

Milind B Ratnaparkhe ◽

...

Keyword(s):

Abiotic Stress ◽

Fatty Acid ◽

Salinity Stress ◽

Stress Responses ◽

Fatty Acid Desaturase ◽

Stress Conditions ◽

Omega 3 ◽

Bean Pod Mottle Virus ◽

Relative Water ◽

Soybean Plants

Abstract Fatty acid desaturases (FADs) are a class of enzymes that mediate desaturation of fatty acids by introducing double bonds. They play an important role in modulating membrane fluidity in response to various abiotic stresses. However, a comprehensive analysis of FAD3 in drought and salinity stress tolerance in soybean is lacking. We used Bean Pod Mottle Virus (BPMV)-based vector for achieving rapid and efficient overexpression as well as silencing of Omega-3 Fatty Acid Desaturase gene from Glycine max (GmFAD3) to assess the functional role of FAD3 in abiotic stress responses in soybean. Higher levels of recombinant BPMV-GmFAD3A transcripts were detected in overexpressing soybean plants. Overexpression of GmFAD3A in soybean resulted in increased levels of jasmonic acid and higher expression of GmWRKY54 as compared to mock-inoculated, vector-infected and FAD3-silenced soybean plants under drought and salinity stress conditions. FAD3A overexpressing plants showed higher levels of chlorophyll content, efficient photosystem-II, relative water content, transpiration rate, stomatal conductance, proline content and also cooler canopy under drought and salinity stress conditions as compared to mock-inoculated, vector-infected and FAD3-silenced soybean plants. Results from the current study revealed that GmFAD3A overexpressing soybean plants exhibited tolerance to drought and salinity stresses. However, soybean plants silenced for GmFAD3 were vulnerable to drought and salinity stresses.

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