Get 20M+ Full-Text Papers For Less Than $1.50/day. Start a 14-Day Trial for You or Your Team.

Learn More →

Preparation and Properties of the Recombinant Tenebrio molitor SerPH122—Proteolytically Active Homolog of Serine Peptidase

Preparation and Properties of the Recombinant Tenebrio molitor SerPH122—Proteolytically Active... Pseudoenzymes are homologs of active enzymes that have amino acid substitutions in the active center and, therefore, usually do not possess enzymatic activity. In this work, a recombinant proprotein (proSerPH122) of the homolog of serine peptidases of the S1 family from the yellow mealworm (Tenebrio molitor) was obtained in the yeast producer strain Komagataella kurtzmanii. The target His6-tagged protein was produced in a glycosylated form during secretion in yeast. The properties of both glycosylated and deglycosylated forms were studied. The proSerPH122 homolog with the replacement of active site Ser with Thr was pretreated with trypsin to study the enzymatic properties. The processed, mature homolog SerPH122 was shown to have low, but reliably detectable activity on the chromogenic substrate Suc-Ala-Ala-Pro-Phe-pNA, and this activity did not depend on the glycosylation level. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Applied Biochemistry and Microbiology Springer Journals

Preparation and Properties of the Recombinant Tenebrio molitor SerPH122—Proteolytically Active Homolog of Serine Peptidase

Loading next page...
 
/lp/springer-journals/preparation-and-properties-of-the-recombinant-tenebrio-molitor-Gds64F0RpV
Publisher
Springer Journals
Copyright
Copyright © Pleiades Publishing, Inc. 2021. ISSN 0003-6838, Applied Biochemistry and Microbiology, 2021, Vol. 57, No. 5, pp. 579–585. © Pleiades Publishing, Inc., 2021. Russian Text © The Author(s), 2021, published in Prikladnaya Biokhimiya i Mikrobiologiya, 2021, Vol. 57, No. 5, pp. 450–457.
ISSN
0003-6838
eISSN
1608-3024
DOI
10.1134/s0003683821050161
Publisher site
See Article on Publisher Site

Abstract

Pseudoenzymes are homologs of active enzymes that have amino acid substitutions in the active center and, therefore, usually do not possess enzymatic activity. In this work, a recombinant proprotein (proSerPH122) of the homolog of serine peptidases of the S1 family from the yellow mealworm (Tenebrio molitor) was obtained in the yeast producer strain Komagataella kurtzmanii. The target His6-tagged protein was produced in a glycosylated form during secretion in yeast. The properties of both glycosylated and deglycosylated forms were studied. The proSerPH122 homolog with the replacement of active site Ser with Thr was pretreated with trypsin to study the enzymatic properties. The processed, mature homolog SerPH122 was shown to have low, but reliably detectable activity on the chromogenic substrate Suc-Ala-Ala-Pro-Phe-pNA, and this activity did not depend on the glycosylation level.

Journal

Applied Biochemistry and MicrobiologySpringer Journals

Published: Sep 1, 2021

Keywords: pseudoenzymes; pseudopeptidases; homolog of serine peptidases of the S1 family; production of a recombinant protein; Tenebrio molitor

References