Development of a novel cellulase biosensor that detects crystalline cellulose hydrolysis using a transcriptional regulator

作者：Kwon Kil Koang; Yeom Soo Jin; Lee Dae Hee; Jeong Ki Jun; Lee Seung Goo^*

来源：Biochemical and Biophysical Research Communications, 2018, 495(1): 1328-1334.

DOI：10.1015/j.bbrc.2017.11.157

• Biosensor
• Genetic circuit
• Transcriptional regulator
• Cellulase
• Crystalline cellulose

Successful utilization of cellulose as renewable biomass depends on the development of economically feasible technologies, which can aid in enzymatic hydrolysis. In this study, we developed a whole-cell biosensor for detecting cellulolytic activity that relies on the recognition of cellobiose using the transcriptional factor Ce1R from Therrnobifida fusca and transcriptional activation of its downstream gfp reporter gene. The fluorescence intensity of whole-cell biosensor, which was named as cellobiosedetectible generic enzyme screening system (CBGESS), was directly proportional to the concentration of cellobiose. The strong fluorescence intensity of CBGESS demonstrated the ability to detect cellulolytic activity with two cellulosic substrates, carboxymethyl cellulose and p-nitrophenyl beta-D-cellobioside in cellulase-expressing Escherichia coli. In addition, CBGESS easily sensed crystalline cellulolytic activity when commercial Celluclast 1.5L was dropped on an Avicel plate. Therefore, CBGESS is a powerful tool for detecting cellulolytic activity with high sensitivity in the presence of soluble or insoluble cellulosic substrates. CBGESS may be further applied to excavate novel cellulases or microbes from both genetic libraries and various environments.

• 出版日期2018-1-1