AIMS: The objective of the study was to produce and characterize the cinnamoyl esterase EstA from the anaerobic fungus Piromyces equi for potential industrial applications. METHODS AND RESULTS: The catalytic domain EstA was produced in Trichoderma reesei. Because the two fungi displayed different genome features, including different codon usage and GC content, a synthetic gene was designed and expressed, leading to the production of the corresponding protein at around 33 mg per litre in the T. reesei culture medium. After the recombinant protein was purified, biochemical characterization showed that EstA presents peak activity at pH 6.5 and at 50-60 degrees C. Furthermore, EstA remained stable at pH 6-8 and below 50 degrees C. EstA was compared to cinnamoyl esterases FaeA and FaeB from Aspergillus niger in terms of ferulic acid (FA) release from wheat bran (WB), maize bran (MB) and sugar beet pulp (SBP). CONCLUSION: The synthetic gene was successfully cloned and overexpressed in T. reesei. EstA from P. equi was demonstrated to efficiently release FA from various natural substrates. SIGNIFICANCE AND IMPACT OF THE STUDY: Recombinant EstA produced in an industrial enzyme producer, T. reesei, was biochemically characterized, and its capacity to release an aromatic compound (FA) for biotechnological applications was demonstrated.
AIMS: The objective of the study was to produce and characterize the cinnamoyl esterase EstA from the anaerobic fungus Piromyces equi for potential industrial applications. METHODS AND RESULTS: The catalytic domain EstA was produced in Trichoderma reesei. Because the two fungi displayed different genome features, including different codon usage and GC content, a synthetic gene was designed and expressed, leading to the production of the corresponding protein at around 33 mg per litre in the T. reesei culture medium. After the recombinant protein was purified, biochemical characterization showed that EstA presents peak activity at pH 6.5 and at 50-60 degrees C. Furthermore, EstA remained stable at pH 6-8 and below 50 degrees C. EstA was compared to cinnamoyl esterases FaeA and FaeB from Aspergillus niger in terms of ferulic acid (FA) release from wheat bran (WB), maize bran (MB) and sugar beet pulp (SBP). CONCLUSION: The synthetic gene was successfully cloned and overexpressed in T. reesei. EstA from P. equi was demonstrated to efficiently release FA from various natural substrates. SIGNIFICANCE AND IMPACT OF THE STUDY: Recombinant EstA produced in an industrial enzyme producer, T. reesei, was biochemically characterized, and its capacity to release an aromatic compound (FA) for biotechnological applications was demonstrated.
Authors: Eunice N Zwane; Petrus J van Zyl; Kwaku G Duodu; Shaunita H Rose; Karl Rumbold; Willem H van Zyl; Marinda Viljoen-Bloom Journal: J Food Sci Technol Date: 2017-02-09 Impact factor: 2.701
Authors: Eunice N Zwane; Shaunita H Rose; Willem H van Zyl; Karl Rumbold; Marinda Viljoen-Bloom Journal: J Ind Microbiol Biotechnol Date: 2014-03-25 Impact factor: 3.346
Authors: Adiphol Dilokpimol; Miia R Mäkelä; Maria Victoria Aguilar-Pontes; Isabelle Benoit-Gelber; Kristiina S Hildén; Ronald P de Vries Journal: Biotechnol Biofuels Date: 2016-10-28 Impact factor: 6.040
Authors: Kerstin Voigt; Timothy Y James; Paul M Kirk; André L C M de A Santiago; Bruce Waldman; Gareth W Griffith; Minjie Fu; Renate Radek; Jürgen F H Strassert; Christian Wurzbacher; Gustavo Henrique Jerônimo; David R Simmons; Kensuke Seto; Eleni Gentekaki; Vedprakash G Hurdeal; Kevin D Hyde; Thuong T T Nguyen; Hyang Burm Lee Journal: Fungal Divers Date: 2021-09-29 Impact factor: 24.902