Production of XynX, a large multimodular protein of thermoanaerobacterium sp., by protease-deficient Bacillus subtilis strains [corrected].
XynX of Thermoanaerobacterium sp. [corrected] is a large, multimodular xylanase of 116 kDa. An Escherichia coli transformant carrying the entire xynX produced three active truncated xylanase species of 105, 85, and 64 kDa intracellularly. The Bacillus subtilis WB700 transformant with the xynX, a strain deficient in seven proteases including Vpr, secreted two active truncated xylanase species of 65 and 44 kDa. The B. subtilis WB800 transformant with xynX, a strain deficient in eight proteases including Vpr and WprA, secreted more active enzymes, 8.46 U ml(-1), mostly in the form of 105 and 85 kDa, than the WB700 transformant, 6.93 U ml(-1). This indicates that the additional deletion of wprA enabled the WB800 to secrete XynX in its intact form. B. subtilis WB800 produced more total enzyme activity than E. coli (1,692 ± 274 U vs. 141.9 ± 27.1 U), and, more importantly, secreted almost all the enzyme activity. The results suggest the potential use of B. subtilis WB800 as a host system for the production of large multimodular proteins.
Department of Agricultural Chemistry, Sunchon National University, Suncheon 540-742, South Korea.
Nguyen Dinh
ND
Phuong
Yu Seok
YS
Jeong
Thangaswamy
T
Selvaraj
Sung Kyum
SK
Kim
Yong Ho
YH
Kim
Kyung Hwa
KH
Jung
Jungho
J
Kim
Han Dae
HD
Yun
Sui-Lam
SL
Wong
Jung-Kul
JK
Lee
Hoon
H
Kim
D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics*; D048013:Acetivibrio thermocellus / Q000201:enzymology* / Q000235:genetics; D043364:Endo-1,4-beta Xylanases / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D010447:Peptide Hydrolases / Q000172:deficiency*
D043364:Endo-1,4-beta Xylanases; D010447:Peptide Hydrolases