Corn bran is the most abundant, low-valued by-product of the industrial corn starch process. It has high amounts of polysaccharide which can be degraded for other use. Thus, developing enzymes for polysaccharide degradation became essential. The transcriptome of Schizophyllum sp. cultured with corn bran fiber as the sole carbon source was determined in this study. A total of 23, 656 UniGenes was obtained. The average GC content of these genes was 58.97%. Sequence analysis showed that there were 5 xylanase genes belonging to the 10th and 11th families of the glycoside hydrolase, and 6 α-L-arabinofuranosidase genes belonging to the 43rd, 51st and 62nd families of glycoside hydrolase. The gene, Sabf32, encoding an α-L-arabinofuranosidase, was isolated, cloned, sequenced, and characterized. It was 975 pb, coding for a protein with 324 amino acid residues and a signal peptide of 21 amino acids. The mature protein had a calculated mass of 32.5 kDa. Characterization of the purified Sabf32 produced in Pichia pastoris revealed its optimum condition being pH at 4.0 and temperature at 40 ℃. The specific enzyme activity of Sabf32 was 16.18 U/mg. This protein was found to be stable at 40 ℃ and under pH 3.5 to 5.5. The evaluated Km and Vmax values with p-nitrophenyl-a-L-arabinofuranoside as substrate were 3.98±0.32 mmol/L and 2.59±0.09 U/mg, respectively. This study provides a basis for further development of corn bran fiber degrading enzymes and synergistic degradation of multiple enzymes.
LIU Yuchun
,
GUO Chao
,
GUO Weiqun
. Transcriptome sequencing of Schizophyllum commune cultured by corn bran fiber fermentation and heterologous expression of recombinant α-L-arabinofuranosidase[J]. Food and Fermentation Industries, 2019
, 45(23)
: 21
-28
.
DOI: 10.13995/j.cnki.11-1802/ts.021903
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