Systematic Screening of All Signal Peptides from Bacillus subtilis: A Powerful Strategy in Optimizing Heterologous Protein Secretion in Gram-positive Bacteria

2006 ◽  
Vol 362 (3) ◽  
pp. 393-402 ◽  
Author(s):  
Ulf Brockmeier ◽  
Michael Caspers ◽  
Roland Freudl ◽  
Alexander Jockwer ◽  
Thomas Noll ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Protein Secretion ◽  
Heterologous Protein ◽  
Signal Peptides ◽  
Gram Positive ◽  
Systematic Screening ◽  
Gram Positive Bacteria ◽  
Heterologous Protein Secretion ◽  
Powerful Strategy

Protein secretion pathways in Bacillus subtilis: Implication for optimization of heterologous protein secretion

2007 ◽  
Vol 25 (1) ◽  
pp. 1-12 ◽  
Author(s):  
Ling Lin Fu ◽  
Zi Rong Xu ◽  
Wei Fen Li ◽  
Jiang Bing Shuai ◽  
Ping Lu ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Protein Secretion ◽  
Heterologous Protein ◽  
Heterologous Protein Secretion ◽  
Secretion Pathways

scholarly journals Heterologous protein secretion byLactobacillus plantarumusing homologous signal peptides

2008 ◽  
Vol 105 (1) ◽  
pp. 215-226 ◽  
Author(s):  
G. Mathiesen ◽  
A. Sveen ◽  
J.-C. Piard ◽  
L. Axelsson ◽  
V.G.H. Eijsink
Keyword(s):  
Protein Secretion ◽  
Heterologous Protein ◽  
Signal Peptides ◽  
Heterologous Protein Secretion

scholarly journals Improvement of Heterologous Protein Secretion by Bacillus subtilis

10.5772/29256 ◽  
2012 ◽  
Author(s):  
Hiroshi Kakeshita ◽  
Yasushi Kageyama ◽  
Katsuya Ozaki ◽  
Kouji Nakamura ◽  
Katsutoshi Ar
Keyword(s):  
Bacillus Subtilis ◽  
Protein Secretion ◽  
Heterologous Protein ◽  
Heterologous Protein Secretion

scholarly journals An optimized growth medium for increased recombinant protein secretion titer via the type III secretion system

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Lisa Ann Burdette ◽  
Han Teng Wong ◽  
Danielle Tullman-Ercek
Keyword(s):  
Protein Secretion ◽  
Salt Concentration ◽  
Type Iii Secretion ◽  
Heterologous Protein ◽  
Carbon Sources ◽  
Type Iii Secretion System ◽  
Secretion System ◽  
Type Iii ◽  
Heterologous Protein Secretion ◽  
Extracellular Environment

Abstract Background Protein secretion in bacteria is an attractive strategy for heterologous protein production because it retains the high titers and tractability of bacterial hosts while simplifying downstream processing. Traditional intracellular production strategies require cell lysis and separation of the protein product from the chemically similar cellular contents, often a multi-step process that can include an expensive refolding step. The type III secretion system of Salmonella enterica Typhimurium transports proteins from the cytoplasm to the extracellular environment in a single step and is thus a promising solution for protein secretion in bacteria. Product titer is sensitive to extracellular environmental conditions, however, and T3SS regulation is integrated with essential cellular functions. Instead of attempting to untangle a complex web of regulatory input, we took an “outside-in” approach to elucidate the effect of growth medium components on secretion titer. Results We dissected the individual and combined effects of carbon sources, buffers, and salts in a rich nutrient base on secretion titer. Carbon sources alone decreased secretion titer, secretion titer increased with salt concentration, and the combination of a carbon source, buffer, and high salt concentration had a synergistic effect on secretion titer. Transcriptional activity measured by flow cytometry showed that medium composition affected secretion system activity, and prolonged secretion system activation correlated strongly with increased secretion titer. We found that an optimal combination of glycerol, phosphate, and sodium chloride provided at least a fourfold increase in secretion titer for a variety of proteins. Further, the increase in secretion titer provided by the optimized medium was additive with strain enhancements. Conclusions We leveraged the sensitivity of the type III secretion system to the extracellular environment to increase heterologous protein secretion titer. Our results suggest that maximizing secretion titer via the type III secretion system is not as simple as maximizing secreted protein expression—one must also optimize secretion system activity. This work advances the type III secretion system as a platform for heterologous protein secretion in bacteria and will form a basis for future engineering efforts.

scholarly journals In Vitro Characterization of the Bacillus subtilis Protein Tyrosine Phosphatase YwqE

2005 ◽  
Vol 187 (10) ◽  
pp. 3384-3390 ◽  
Author(s):  
Ivan Mijakovic ◽  
Lucia Musumeci ◽  
Lutz Tautz ◽  
Dina Petranovic ◽  
Robert A. Edwards ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Tyrosine Phosphatase ◽  
Protein Tyrosine Phosphatases ◽  
Class Ii ◽  
Gram Positive ◽  
Protein Tyrosine ◽  
Gram Positive Bacteria ◽  
New Family ◽  
In Vitro Characterization

ABSTRACT Both gram-negative and gram-positive bacteria possess protein tyrosine phosphatases (PTPs) with a catalytic Cys residue. In addition, many gram-positive bacteria have acquired a new family of PTPs, whose first characterized member was CpsB from Streptococcus pneumoniae. Bacillus subtilis contains one such CpsB-like PTP, YwqE, in addition to two class II Cys-based PTPs, YwlE and YfkJ. The substrates for both YwlE and YfkJ are presently unknown, while YwqE was shown to dephosphorylate two phosphotyrosine-containing proteins implicated in UDP-glucuronate biosynthesis, YwqD and YwqF. In this study, we characterize YwqE, compare the activities of the three B. subtilis PTPs (YwqE, YwlE, and YfkJ), and demonstrate that the two B. subtilis class II PTPs do not dephosphorylate the physiological substrates of YwqE.

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