Establishing regiocontrol of disulfide bond isomers of α-conotoxin ImI via the synthesis of N-to-C cyclic analogs

Biopolymers ◽  
2009 ◽  
Vol 94 (3) ◽  
pp. 307-313 ◽  
Author(s):  
Christopher J. Armishaw ◽  
Julie L. Dutton ◽  
David J. Craik ◽  
Paul F. Alewood
Keyword(s):  
Disulfide Bond ◽  
Bond Isomers

Bombyxin-II and its disulfide bond isomers: Synthesis and activity

Peptides ◽  
1992 ◽  
Vol 13 (4) ◽  
pp. 653-662 ◽  
Author(s):  
Koji Nagata ◽  
Kazunori Maruyama ◽  
Hiromichi Nagasawa ◽  
Ikumi Urushibata ◽  
Akira Isogai ◽  
...  
Keyword(s):  
Disulfide Bond ◽  
Bond Isomers

Disulfide Bond Structure and Accessibility of Cysteines in the Ectodomain of the Cholecystokinin Receptor: Specific Mono-Reactive Receptor Constructs Examine Charge-Sensitivity of Loop Regions

2003 ◽  
Vol 9 (2) ◽  
pp. 83-91
Author(s):  
Xi-Qin Ding ◽  
Vesile Dolu ◽  
Elizabeth M. Hadac ◽  
Michael Schuetz ◽  
Laurence J. Miller
Keyword(s):  
Disulfide Bond ◽  
Cholecystokinin Receptor ◽  
Loop Regions

Van der Waals interactions of the disulfide bond revealed: A microwave spectroscopic study of the diethyl disulfide–argon complex

2021 ◽  
Vol 154 (12) ◽  
pp. 124306
Author(s):  
Tao Lu ◽  
Daniel A. Obenchain ◽  
Jiaqi Zhang ◽  
Jens-Uwe Grabow ◽  
Gang Feng
Keyword(s):  
Spectroscopic Study ◽  
Disulfide Bond ◽  
Van Der Waals ◽  
Van Der Waals Interactions

scholarly journals Disulfide bond engineering of AppA phytase for increased thermostability requires co-expression of protein disulfide isomerase in Pichia pastoris

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Laura Navone ◽  
Thomas Vogl ◽  
Pawarisa Luangthongkam ◽  
Jo-Anne Blinco ◽  
Carlos H. Luna-Flores ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Phytic Acid ◽  
Disulfide Bond ◽  
Wild Type ◽  
Wild Type Enzyme ◽  
E Coli ◽  
Biochemical Characterisation ◽  
Disulfide Bond Isomerase ◽  
Microbial Systems ◽  
Protein Disulfide

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.

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