Skip to main content
SpringerLink
Log in

Cloning and sequence analysis of putative type II fatty acid synthase genes from Arachis hypogaea L.

  • Published:
Journal of Biosciences Aims and scope Submit manuscript

Abstract

The cultivated peanut is a valuable source of dietary oil and ranks fifth among the world oil crops. Plant fatty acid biosynthesis is catalysed by type II fatty acid synthase (FAS) in plastids and mitochondria. By constructing a full-length cDNA library derived from immature peanut seeds and homology-based cloning, candidate genes of acyl carrier protein (ACP), malonyl-CoA:ACP transacylase, β-ketoacyl-ACP synthase (I, II, III), β-ketoacyl-ACP reductase, β-hydroxyacyl-ACP dehydrase and enoyl-ACP reductase were isolated. Sequence alignments revealed that primary structures of type II FAS enzymes were highly conserved in higher plants and the catalytic residues were strictly conserved in Escherichia coli and higher plants. Homologue numbers of each type II FAS gene expressing in developing peanut seeds varied from 1 in KASII, KASIII and HD to 5 in ENR. The number of single-nucleotide polymorphisms (SNPs) was quite different in each gene. Peanut type II FAS genes were predicted to target plastids except ACP2 and ACP3. The results suggested that peanut may contain two type II FAS systems in plastids and mitochondria. The type II FAS enzymes in higher plants may have similar functions as those in E. coli.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

ACAT:

acetyl CoA:ACP transacylase

ACP:

acyl carrier protein

DAP:

days after pegging

ENR:

enoyl-ACP reductase

EST:

expressed sequence tag

FAS:

fatty acid synthase

HD:

β-hydroxyacyl-ACP dehydrase

KAS:

β-ketoacyl-ACP synthase

KR:

β-ketoacyl-ACP reductase

MCAT:

malonyl-CoA:ACP transacylase

NCBI:

National Center for Biotechnology

ORF:

open reading frame

PCR:

polymerase chain reaction

RACE:

rapid amplification of cDNA ends

SNP:

single-nucleotide polymorphism

References

  • Abbadi A, Brummel M and Spener F 2000 Knockout of the regulatory site of 3-ketoacyl-ACP synthase III enhances shortand medium-chain acyl-ACP synthesis; Plant J. 24 1–9

    Article  PubMed  CAS  Google Scholar 

  • Bolle C, Herrmann R G and Oelmuller R 1996 Different sequences for 5′-untranslated leaders of nuclear genes for plastid proteins affect the expression of the beta-glucuronidase gene; Plant Mol. Biol. 32 861–868

    Article  PubMed  CAS  Google Scholar 

  • Bonaventure G and Ohlrogge J B 2002 Differential regulation of mRNA levels of acyl carrier protein isoforms in Arabidopsis; Plant Physiol. 128 223–235

    Article  PubMed  CAS  Google Scholar 

  • Carlsson A S, LaBrie S T, Kinney A J, von Wettstein-Knowles P and Browse J 2002 A KAS2 cDNA complements the phenotypes of the Arabidopsis fab1 mutant that differs in a single residue bordering the substrate binding pocket; Plant J. 29 761–770

    Article  PubMed  CAS  Google Scholar 

  • Chen J and Post-Beittenmiller D 1996 Molecular cloning of a cDNA encoding 3-ketoacyl-acyl carrier protein synthase III from leek; Gene 182 45–52

    Article  PubMed  CAS  Google Scholar 

  • Clough R C, Matthis A L, Barnum S R and Jaworski J G 1992 Purification and characterization of 3-ketoacyl-acyl carrier protein synthase III from spinach; J. Biol. Chem. 267 20992–20998

    PubMed  CAS  Google Scholar 

  • Dehesh K, Tai H, Edwards P, Byrne J and Jaworski J G 2001 Overexpression of 3-ketoacyl-acyl-carrier protein synthase IIIs in plants reduces the rate of lipid synthesis; Plant Physiol. 125 1103–1114

    Article  PubMed  CAS  Google Scholar 

  • Fisher M, Kroon J T M, Martindale W, Stuitje A R, Slabas A R and Rafferty J B 2000 The X-ray structure of Brassica napus β-keto acyl carrier protein reductase and its implications for substrate binding and catalysis; Structure 8 339–347

    Article  PubMed  CAS  Google Scholar 

  • Gulliver B S and Slabas A R 1994 Acetoacyl-acyl carrier protein synthase from avocado: its purification, characterisation and clear resolution from acetyl CoA:ACP transacylase; Plant Mol. Biol. 25 179–191

    Article  PubMed  CAS  Google Scholar 

  • Han L, Lobo S and Reynold K A 1998 Characterization of β-ketoacyl-acyl carrier protein synthase III from Streptomyces glaucescens and its role in initiation of fatty acid biosynthesis; J. Bacteriol. 180 4481–4486

    PubMed  CAS  Google Scholar 

  • Hansen L and Kauppinen S 1991 Barley acyl carrier protein II: nucleotide sequence of cDNA clones and chromosomal location of the Acl2 gene; Plant Physiol. 97 472-474

    Google Scholar 

  • Hansen and von Wettstein-Knowles P 1991 The barley genes Acl1 and Acl3 encoding acyl carrier proteins I and III are located on different chromosomes; Mol. Gen. Genet. 229 467–478

    Article  PubMed  CAS  Google Scholar 

  • Heath R J and Rock C O 1996 Roles of the FabA and FabZ β-hydroxyacyl-acyl carrier protein dehydratases in Escherichia coli fatty acid biosynthesis; J. Biol. Chem. 271 27795–27801

    Article  PubMed  CAS  Google Scholar 

  • Hlousek-Radojcic A, Post-Beittenmiller D and Ohlrogge JB 1992 Expression of constitutive and tissue-specific acyl carrier protein isoforms in Arabidopsis; Plant Physiol. 98 206–214

    Article  PubMed  CAS  Google Scholar 

  • Kater M M, Koningstein G M, Nijkamp H J and Stuitje A R 1991 cDNA cloning and expression of Brassica napus enoyl-acyl carrier protein reductase in Escherichia coli; Plant Mol. Biol. 17 895–909

    Article  PubMed  CAS  Google Scholar 

  • Keatinge-Clay A T, Shelat A A, Savage D F, Tsai S C, Miercke L J, O’Connell J D, Khosla C and Stroud R M 2003 Catalysis, specificity, and ACP docking site of Streptomyces coelicolor malonyl-CoA:ACP transacylase; Structure 11 147–154

    Article  PubMed  CAS  Google Scholar 

  • Kimber M S, Martin F, Lu Y J, Houston S, Vedadi M, Dharamsi A, Fiebig K M, Schmid M and Rock C O 2004 The structure of (3R)-hydroxyacyl-acyl carrier protein dehydratase (FabZ) from Pseudomonas aeruginosa; J. Biol. Chem. 2792 52593–52602

    Article  Google Scholar 

  • Klein B, Pawlowski K, Horicke-Grandpierre C, Schell J and Topfer R 1992 Isolation and characterization of a cDNA from Cuphea lanceolata encoding a beta-ketoacyl-ACP reductase; Mol. Gen. Genet. 233 122–128

    Article  PubMed  CAS  Google Scholar 

  • Kopka J, Robers M, Schuch R and Spener F 1993 Acyl carrier proteins from developing seeds of Cuphea lanceolata Ait; Planta 191 102–111

    Article  CAS  Google Scholar 

  • Lai C Y and Cronan J E 2003 β-ketoacyl-acyl carrier protein synthase III (FabH) is essential for bacterial fatty acid synthesis; J. Biol. Chem. 278 51494–51503

    Article  PubMed  CAS  Google Scholar 

  • Lamppa G and Jacks C 1991 Analysis of two linked genes coding for the acyl carrier protein (ACP) from Arabidopsis thaliana (columbia); Plant Mol. Biol. 16 469–474

    Article  PubMed  CAS  Google Scholar 

  • Leesong M, Henderson B S, Gillig J R, Schwab J M and Smith J L 1996 Structure of a dehydratase-isomerase from the bacterial pathway for biosynthesis of unsaturated fatty acids: two catalytic activities in one active site; Structure 4 253–264

    Article  PubMed  CAS  Google Scholar 

  • Mekhedov S, Cahoon E B and Ohlrogge J 2001 An unusual seed-specific 3-ketoacyl-ACP synthase associated with the biosynthesis of petroselinic acid in coriander; Plant Mol. Biol. 47 507–518

    Article  PubMed  CAS  Google Scholar 

  • Mofid M R, Finking R and Marahiel M A 2002 Recognition of hybrid peptidyl carrier proteins/acyl carrier proteins in nonribosomal peptide synthetase modules by the 4′-phosphopan tetheinyl transferases AcpS and Sfp; J. Biol. Chem. 277 17023–17031

    Article  PubMed  CAS  Google Scholar 

  • Moretzsohn M C, Hopkins M S, Mitchell S E, Kresovich S, Valls J F M and Ferreira M E 2004 Genetic diversity of peanut (Arachis hypogaea L.) and its wild relatives based on the analysis of hypervariable regions of the genome; BMC Plant Biol. 4 11

    Article  Google Scholar 

  • Olsen J G, Kadziola A, von Wettstein-Knowles P, Siggaard-Andersen M, Lindquist Y and Larsen S 1999 The X-ray crystal structure of beta-ketoacyl [acyl carrier protein] synthase I; FEBS Lett. 460 46–52

    Article  PubMed  CAS  Google Scholar 

  • Olsen J G, Rasmussen A V, von Wettstein-Knowles P and Henriksen A 2004 Structure of the mitochondrial beta-ketoacyl-[acyl carrier protein] synthase from Arabidopsis and its role in fatty acid synthesis; FEBS Lett. 577 170–174

    Article  PubMed  CAS  Google Scholar 

  • Pidkowich M S, Nguyen H T, Heilmann I, Ischebeck T and Shanklin J 2007 Modulating seed beta-ketoacyl-acyl carrier protein synthase II level converts the composition of a temperate seed oil to that of a palm-like tropical oil;. Proc. Natl. Acad. Sci USA 104 4742–4747

    Article  PubMed  CAS  Google Scholar 

  • Post-Beittenmiller M A, Hlousek-Radojcic A and Ohlrogge J B 1989 DNA sequence of a genomic clone encoding an Arabidopsis acyl carrier protein (ACP); Nucleic Acids Res. 17 1777

    Article  PubMed  CAS  Google Scholar 

  • Price A C, Zhang Y M, Rock C O and White S W 2001 Structure of β-ketoacyl-[acyl carrier protein] reductase from Escherichia coli: negative cooperativity and its structural basis; Biochemistry 40 12772–12781

    Article  PubMed  CAS  Google Scholar 

  • Qiu X, Janson C A, Konstantinidis A K, Nwagwu S, Silverman C, Smith W W, Khandekar S, Lonsdale J and Abdel-Meguid S S 1999 Crystal structure of beta-ketoacyl-acyl carrier protein synthase III. A key condensing enzyme in bacterial fatty acid biosynthesis; J. Biol. Chem. 274 36465–36471

    Article  PubMed  CAS  Google Scholar 

  • Rafferty J B, Simon J W, Baldock C, Artymiuk P J, Baker P J, Stuitje A R, Slabas A R and Rice D W 1995 Common themes in redox chemistry emerge from the X-ray structure of oilseed rape (Brassica napus) enoyl acyl carrier protein reductase; Structure 3 927–938

    Article  PubMed  CAS  Google Scholar 

  • Rafi S, Novichenok P, Kolappan S, Zhang X, Stratton C F, Rawat R, Kisker C, Simmerling C and Tonge P J 2006 Structure of acyl carrier protein bound to FabI, the FASII enoyl reductase from Escherichia coli; J. Biol. Chem. 281 39285–39293

    Article  PubMed  CAS  Google Scholar 

  • Scherer D E and Knauf V C 1987 Isolation of a cDNA clone for the acyl carrier protein-I of spinach; Plant Mol. Biol. 9 127–134

    Article  CAS  Google Scholar 

  • Schmid K M and Ohlrogge J B 1990 A root acyl carrier protein-II from spinach is also expressed in leaves and seeds; Plant Mol. Biol. 15 765–778

    Article  PubMed  CAS  Google Scholar 

  • Serre L, Verbree E C, Dauter Z, Stuitje A R and Derewenda Z S 1995 The Escherichia coli malonyl-CoA:acyl carrier protein transacylase at 1.5-A resolution. Crystal structure of a fatty acid synthase component; J. Biol. Chem. 270 12961–12964

    Article  PubMed  CAS  Google Scholar 

  • Shintani D K and Ohlrogge J B 1994 The characterization of a mitochondrial acyl carrier protein isoform isolated from Arabidopsis thaliana; Plant Physiol. 104 1221–1229

    Article  PubMed  CAS  Google Scholar 

  • Simon J W and Slabas A R 1998 cDNA cloning of Brassica napus malonyl-CoA:ACP transacylase (MCAT) (fabD) and complementation of an E. coli MCAT mutant; FEBS Lett. 435 204–206

    Article  PubMed  CAS  Google Scholar 

  • Slabaugh M B, Tai H, Jaworski J and Knapp S J 1995 cDNA clones encoding beta-ketoacyl-acyl carrier protein synthase III from Cuphea wrightii; Plant Physiol. 108 443–444

    Article  PubMed  CAS  Google Scholar 

  • Suh M C, Schultz D J and Ohlrogge J B 1999 Isoforms of acyl carrier protein involved in seed-specific fatty acid synthesis; Plant J. 17 679–688

    Article  PubMed  CAS  Google Scholar 

  • Tai H and Jaworski J G 1993 3-Ketoacyl-acyl carrier protein synthase III from spinach (Spinacia oleracea) is not similar to other condensing enzymes of fatty acid synthase; Plant Physiol. 103 1361–1367

    Article  PubMed  CAS  Google Scholar 

  • Tai H, Post-Beittenmiller D and Jaworski J G 1994 Cloning of a cDNA encoding 3-ketoacyl-acyl carrier protein synthase III from Arabidopsis; Plant Physiol. 106 801-802

    Google Scholar 

  • Tsay J T, Oh W, Larson T J, Jackowski S and Rock C O 1992 Isolation and characterization of the β-ketoacyl-acyl carrier protein synthase III gene (fabH) from Escherichia coli K-12; J. Biol. Chem. 267 6807–6814

    PubMed  CAS  Google Scholar 

  • Voetz M, Klein B, Schell J and Topfer R 1994 Three different cDNAs encoding acyl carrier proteins from Cuphea lanceolata; Plant Physiol. 106 785–786

    Article  PubMed  CAS  Google Scholar 

  • von Wettstein-Knowles P, Olsen J G, McGuire K A and Larsen S 2000 Molecular aspects of β-ketoacyl synthase (KAS) catalysis; Biochem. Soc. Trans. 28 601–607

    Article  Google Scholar 

  • Wada H, Shintani D K and Ohlrogge J B 1997 Why do mitochondria synthesize fatty acids? Evidence for involvement in lipoic acid production; Proc. Natl. Acad. Sci. USA 94 1591–1596

    Article  PubMed  CAS  Google Scholar 

  • White S W, Zheng J, Zhang Y M and Rock C O 2005 The structural biology of type II fatty acid biosynthesis; Annu. Rev. Biochem. 74 791–831

    Article  PubMed  CAS  Google Scholar 

  • Yasuno R, von Wettstein-Knowles P and Wada H 2004 Identification and molecular characterization of the beta-ketoacyl-[acyl carrier protein] synthase component of the Arabidopsis mitochondrial fatty acid synthase; J. Biol. Chem. 279 8242–8251

    Article  PubMed  CAS  Google Scholar 

  • Zhang Y M, Marrakchi H, White S W and Rock C O 2003a The application of computational methods to explore the diversity and structure of bacterial fatty acid synthase; J. Lipid Res. 44 1–10

    Article  PubMed  Google Scholar 

  • Zhang Y M, Rao M S, Heath R J, Price A C, Olson A J, Rock C O and White S W 2001 Identification and analysis of the acyl carrier protein (ACP) docking site on β-ketoacyl-ACP synthase III; J. Biol. Chem. 276 8231–8238

    Article  PubMed  CAS  Google Scholar 

  • Zhang Y M, Wu B, Zheng J and Rock C O 2003b Key residues responsible for acyl carrier protein and β-ketoacyl-acyl carrier protein reductase (FabG) interaction; J. Biol. Chem. 278 52935–52943

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. High-Tech Research Center, Shandong Academy of Agricultural Sciences, Key Laboratory for Genetic Improvement of Crop, Animal and Poultry of Shandong Province, Key Laboratory of Crop Genetic Improvement and Biotechnology, Huanghuaihai, Ministry of Agriculture, Ji’nan, 250100, China

    Meng-Jun Li, Ai-Qin Li, Han Xia, Chuan-Zhi Zhao, Chang-Sheng Li, Shu-Bo Wan, Yu-Ping Bi & Xing-Jun Wang

Authors
  1. Meng-Jun Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ai-Qin Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Han Xia
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Chuan-Zhi Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Chang-Sheng Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Shu-Bo Wan
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yu-Ping Bi
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Xing-Jun Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Xing-Jun Wang.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, MJ., Li, AQ., Xia, H. et al. Cloning and sequence analysis of putative type II fatty acid synthase genes from Arachis hypogaea L.. J Biosci 34, 227–238 (2009). https://doi.org/10.1007/s12038-009-0027-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12038-009-0027-1

Keywords

Search

Navigation