title: Purification and characterization of fructan and fructansucrase from Lactobacillus fermentum AKJ15 isolated from Kodo ko jaanr, a fermented beverage from north-eastern Himalayas.
abstract: Fructansucrase and fructan produced from Lactobacillus fermentum AKJ15 were isolated from seeds of Kodo ko jaanr, a fermented mild-alcoholic beverage prepared in North East India. The strain was identified by 16S rRNA gene sequence analysis and biochemical characterization. The strain displayed maximum fructansucrase activity of 4.3 U/ml (1.02 U/mg) at 28°C at 180 rpm. The enzyme purified by polyethylene glycol-400 gave specific activity of 5 U/mg and showed 90 kDa band on non-denaturing Sodium Dodecyl Sulphate-Poly Acrylamide Gel Electrophoresis (SDS-PAGE). The purified enzyme confirmed the presence of fructan by periodic acid Schiff's staining which showed magenta colour bands with both sucrose and raffinose. The strain produced 10.2 mg/ml fructan in broth under optimized culture conditions. The purified fructansucrase displayed V(max) of 5.42 U/mg and K(m) of 16.65 mM. The enzyme showed maximum activity at 30°C and at pH 5. The structure of fructan was analysed by (1)H and (13)C NMR spectra confirming β-(2-1) and β-(2-6) linkages.
authors.affiliation: Department of Biotechnology, Indian Institute of Technology Guwahati, Guwahati 781 039, Assam, India.
authors.forename: Arijita
authors.initials: A
authors.lastname: Dutta
authors.forename: Deeplina
authors.initials: D
authors.lastname: Das
authors.forename: Arun
authors.initials: A
authors.lastname: Goyal
mesh_terms: D001483:Base Sequence; D001628:Beverages / Q000382:microbiology*; D004591:Electrophoresis, Polyacrylamide Gel; D031748:Eleusine / Q000737:chemistry* / Q000382:microbiology; D005285:Fermentation; D005630:Fructans / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D007194:India; D052197:Limosilactobacillus fermentum / Q000235:genetics / Q000378:metabolism*; D015394:Molecular Structure; D008970:Molecular Weight; D028321:Plant Preparations / Q000378:metabolism*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D011887:Raffinose / Q000032:analysis; D012639:Seeds / Q000737:chemistry / Q000382:microbiology; D013045:Species Specificity; D013393:Sucrase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D013395:Sucrose / Q000032:analysis
chemical_list: D005630:Fructans; D028321:Plant Preparations; D012336:RNA, Ribosomal, 16S; D013395:Sucrose; D013393:Sucrase; D011887:Raffinose