Cloning and Characterization of groESL Operon in Acetobacter aceti

Abstract

The groESL operon of Acetobacter aceti was cloned and sequenced. We observed that GroES and GroEL of A. aceti had high amino acid sequence homologies to GroES and GroEL of Escherichia coli and Bacillus subtilis. The upstream region of the groESL operon contained the heat-shock promoter, which was previously reported in alpha-purple proteobacteria, and the highly conserved inverted repeat sequence. Phylogenetic analysis revealed that the A. aceti GroES and GroEL are very closely related to those of other alpha-purple proteobacteria. Transcription of this operon in A. aceti was induced by heat shock as well as by exposure to ethanol and acetic acid, which are present during fermentation of acetic acid. A. aceti that overexpressed the groESL was more resistant than the control strain to Stressors such as heat, ethanol, or acetic acid, indicating that GroES and GroEL are closely associated with the characteristic nature of Acetobacter and play an important role in acetic acid fermentation.

An OmpA Family Protein, a Target of the GinI/GinR Quorum-Sensing System in Gluconacetobacter intermedius , Controls Acetic Acid Fermentation

“…Total RNA was isolated by the hot-phenol method (20) from cells grown at 30°C for 6,8,10,12,16, and 24 h in YPG medium containing 2% (vol/vol) ethanol and 1% (vol/vol) cellulase in a shaking flask. A 0.4-kb fragment containing part of gmpA was amplified by PCR with primers AF and AR (Table 1) using the 3.4-kb SmaI fragment as the template.…”

“…Total RNA was isolated by the hot-phenol method (20) from cells grown at 30°C for 8 h in YPG medium containing 2% (vol/vol) ethanol and 1% (vol/vol) cellulase in a shaking flask. RT was performed using the total RNA with SuperScript III reverse transcriptase (Invitrogen).…”

Proteome analysis of Acetobacter pasteurianus during acetic acid fermentation

“…The heat shock systems DnaKJ and GroELS are general stress chaperones that prevent misfolding and promote the refolding and proper assembly of denatured polypeptides generated under stress conditions. These two systems are known to be induced in different microorganisms by various stresses that result from environmental changes [43,44,51,52]. However, the induction of the different stress proteins is not the same in all microorganisms.…”

Protein Profile of Acetobacter pasteurianus HSZ3-21

“…In this study, HSPs and HSPs-related protein, including DnaK (Hsp70), HtpG (Hsp90), GroEL (Hsp60), HspA (Hsp20/alpha), Clp protease ATP-binding subunit ClpB, ATP-dependent Clp protease proteolytic subunit, and endopeptidase DegP (spot nos. 9,10,14,16,24,26,27,29,30,233,3,207,40,90) were identified, indicating that HSPs may be correlated with the resistance against various stresses such as heat, ethanol, or acetic acid [15,16].…”