Identification of disulfide bond isomerase substrates reveals bacterial virulence factors

Abstract Background Phytases are widely used commercially as dietary supplements for swine and poultry to increase the digestibility of phytic acid. Enzyme development has focused on increasing thermostability to withstand the high temperatures during industrial steam pelleting. Increasing thermostability often reduces activity at gut temperatures and there remains a demand for improved phyases for a growing market. Results In this work, we present a thermostable variant of the E. coli AppA phytase, ApV1, that contains an extra non-consecutive disulfide bond. Detailed biochemical characterisation of ApV1 showed similar activity to the wild type, with no statistical differences in kcat and KM for phytic acid or in the pH and temperature activity optima. Yet, it retained approximately 50% activity after incubations for 20 min at 65, 75 and 85 °C compared to almost full inactivation of the wild-type enzyme. Production of ApV1 in Pichia pastoris (Komagataella phaffi) was much lower than the wild-type enzyme due to the presence of the extra non-consecutive disulfide bond. Production bottlenecks were explored using bidirectional promoters for co-expression of folding chaperones. Co-expression of protein disulfide bond isomerase (Pdi) increased production of ApV1 by ~ 12-fold compared to expression without this folding catalyst and restored yields to similar levels seen with the wild-type enzyme. Conclusions Overall, the results show that protein engineering for enhanced enzymatic properties like thermostability may result in folding complexity and decreased production in microbial systems. Hence parallel development of improved production strains is imperative to achieve the desirable levels of recombinant protein for industrial processes.

Download Full-text

Interference of quorum sensing regulated bacterial virulence factors and biofilms by Plumbago zeylanica extract

Microscopy Research and Technique ◽

10.1002/jemt.23872 ◽

2021 ◽

Author(s):

Faizan Abul Qais ◽

Iqbal Ahmad ◽

Fohad Mabood Husain ◽

Suliman Y. Alomar ◽

Naushad Ahmad ◽

...

Keyword(s):

Quorum Sensing ◽

Virulence Factors ◽

Bacterial Virulence ◽

Plumbago Zeylanica

Download Full-text

Host and Bacterial Virulence Factors Predisposing to Emphysematous Pyelonephritis

American Journal of Kidney Diseases ◽

10.1053/j.ajkd.2005.05.019 ◽

2005 ◽

Vol 46 (3) ◽

pp. 432-439 ◽

Cited By ~ 39

Author(s):

Chin-Chung Tseng ◽

Jiunn-Jong Wu ◽

Ming-Cheng Wang ◽

Lien-I. Hor ◽

Yen-Hsiu Ko ◽

...

Keyword(s):

Virulence Factors ◽

Bacterial Virulence ◽

Emphysematous Pyelonephritis

Download Full-text

The role of disulfide bond isomerase A (DsbA) ofEscherichia coliO157:H7 in biofilm formation and virulence

FEMS Microbiology Letters ◽

10.1111/j.1574-6968.2007.00993.x ◽

2008 ◽

Vol 278 (2) ◽

pp. 213-222 ◽

Cited By ~ 26

Author(s):

Yunho Lee ◽

Younghoon Kim ◽

Sujin Yeom ◽

Saehun Kim ◽

Sungsu Park ◽

...

Keyword(s):

Disulfide Bond ◽

Biofilm Formation ◽

Disulfide Bond Isomerase

Download Full-text

Inhibition of bacterial virulence factors of foodborne pathogens by paprika (Capsicum annuum L.) extracts

Food Control ◽

10.1016/j.foodcont.2021.108568 ◽

2021 ◽

pp. 108568

Author(s):

Rocío Daniela Inés Molina ◽

Rodrigo Campos-Silva ◽

Myriam Anabel Díaz ◽

Alexandre José Macedo ◽

María Amparo Blázquez ◽

...

Keyword(s):

Capsicum Annuum ◽

Virulence Factors ◽

Foodborne Pathogens ◽

Bacterial Virulence ◽

Capsicum Annuum L

Download Full-text

Role of Bacterial Virulence Factors and Host Factors in the Outcome of Escherichia coli Bacteraemia

European Journal of Clinical Microbiology & Infectious Diseases ◽

10.1007/s100960050483 ◽

2000 ◽

Vol 19 (4) ◽

pp. 312-316 ◽

Cited By ~ 19

Author(s):

T. A. M. Hekker ◽

A. B. J. Groeneveld ◽

A. M. Simoons-Smit ◽

P. de Man ◽

H. Connell ◽

...

Keyword(s):

Escherichia Coli ◽

Virulence Factors ◽

Bacterial Virulence ◽

Host Factors

Download Full-text

Liposomes as novel anti-infectives targeting bacterial virulence factors?

Expert Review of Anti-infective Therapy ◽

10.1586/14787210.2015.1028367 ◽

2015 ◽

Vol 13 (5) ◽

pp. 531-533 ◽

Cited By ~ 11

Author(s):

Samareh Azeredo da Silveira ◽

Antonio Perez

Keyword(s):

Virulence Factors ◽

Bacterial Virulence

Download Full-text

The Pseudomonas syringae avrRpt2 Gene Contributes to Virulence on Tomato

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi-18-0626 ◽

2005 ◽

Vol 18 (7) ◽

pp. 626-633 ◽

Cited By ~ 27

Author(s):

Melisa T. S. Lim ◽

Barbara N. Kunkel

Keyword(s):

Pseudomonas Syringae ◽

Virulence Factors ◽

Host Defense ◽

Virulence Gene ◽

Bacterial Virulence ◽

Host Tissue ◽

Native Strain ◽

Phytopathogenic Bacteria ◽

Gram Negative ◽

Mutant Derivative

In order to cause disease on plants, gram-negative phytopathogenic bacteria introduce numerous virulence factors into the host cell in order to render host tissue more hospitable for pathogen proliferation. The mode of action of such bacterial virulence factors and their interaction with host defense pathways remain poorly understood. avrRpt2, a gene from Pseudomonas syringae pv. tomato JL1065, has been shown to promote the virulence of heterologous P. syringae strains on Arabidopsis thaliana. However, the contribution of avrRpt2 to the virulence of JL1065 has not been examined previously. We show that a mutant derivative of JL1065 that carries a disruption in avrRpt2 is impaired in its ability to cause disease on tomato (Lycopersicon esculentum), indicating that avrRpt2 also acts as a virulence gene in its native strain on a natural host. The virulence activity of avrRpt2 was detectable on tomato lines that are defective in either ethylene perception or the accumulation of salicylic acid, but could not be detected on a tomato mutant insensitive to jasmonic acid. The enhanced virulence conferred by the expression of avrRpt2 in JL1065 was not associated with the suppression of several defense-related genes induced during the infection of tomato.

Download Full-text