title: Isolation of eicosapentaenoic acid-producing fungi from soil based on polymerase chain reaction amplification.
abstract: A method was developed for rapid screening and isolation of eicosapentaenoic acid (EPA)-producing soil fungi through polymerase chain reaction (PCR) amplification. Genes coding for delta6 fatty acid desaturase and delta5 fatty acid desaturase were used as molecular markers for screening these EPA-producing fungi from soil. Three out of 65 soil fungi gave positive results through PCR amplification. Two out of these three strains were found to produce EPA when they had grown in 80 ml potato/dextrose liquid medium at (25 +/- 1) degrees C for 144 h. The EPA yields were 215.81 mg 1(-1) and 263.80 mg 1(-1), respectively. The other positive strain was detected to produce arachidonic acid (AA). This study indicates that molecular detection of genes encoding delta6 and delta5 desaturases is an efficient method for primary screening of EPA- or its related polyunsaturated fatty acids (PuFAs)-producing fungi, which can improve the screening efficiency prominently.
authors.affiliation: Key Laboratory of Molecular Biophysics, Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China.
authors.forename: Jing-Rong
authors.initials: JR
authors.lastname: Hu
authors.forename: Peng-Peng
authors.initials: PP
authors.lastname: Zhou
authors.forename: Yuan-Min
authors.initials: YM
authors.lastname: Zhu
authors.forename: Liang
authors.initials: L
authors.lastname: Ren
authors.forename: Long-Jiang
authors.initials: LJ
authors.lastname: Yu
mesh_terms: D001483:Base Sequence; D017931:DNA Primers; D015118:Eicosapentaenoic Acid / Q000096:biosynthesis*; D005658:Fungi / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D016133:Polymerase Chain Reaction / Q000379:methods*; D012988:Soil Microbiology
chemical_list: D017931:DNA Primers; D015118:Eicosapentaenoic Acid