Characterization of a novel cold-adapted intracellular serine protease from the extremophile Planococcus halocryophilus Or1

Casper Bøjer Rasmussen; Carsten Scavenius; Ida B Thøgersen; Seandean Lykke Harwood; Øivind Larsen; Gro Elin Kjaereng Bjerga; Peter Stougaard; Jan J Enghild; Mariane Schmidt Thøgersen

doi:10.3389/fmicb.2023.1121857

Characterization of a novel cold-adapted intracellular serine protease from the extremophile Planococcus halocryophilus Or1

Casper Bøjer Rasmussen, Carsten Scavenius, Ida B Thøgersen, Seandean Lykke Harwood, Øivind Larsen, Gro Elin Kjaereng Bjerga, Peter Stougaard, Jan J Enghild, Mariane Schmidt Thøgersen^*

^*Corresponding author for this work

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Abstract

The enzymes of microorganisms that live in cold environments must be able to function at ambient temperatures. Cold-adapted enzymes generally have less ordered structures that convey a higher catalytic rate, but at the cost of lower thermodynamic stability. In this study, we characterized P355, a novel intracellular subtilisin protease (ISP) derived from the genome of Planococcus halocryophilus Or1, which is a bacterium metabolically active down to -25°C. P355's stability and activity at varying pH values, temperatures, and salt concentrations, as well as its temperature-dependent kinetics, were determined and compared to an uncharacterized thermophilic ISP (T0099) from Parageobacillus thermoglucosidasius, a previously characterized ISP (T0034) from Planococcus sp. AW02J18, and Subtilisin Carlsberg (SC). The results showed that P355 was the most heat-labile of these enzymes, closely followed by T0034. P355 and T0034 exhibited catalytic constants (k cat ) that were much higher than those of T0099 and SC. Thus, both P355 and T0034 demonstrate the characteristics of the stability-activity trade-off that has been widely observed in cold-adapted proteases.

Original language	English
Article number	1121857
Journal	Frontiers in Microbiology
Volume	14
Number of pages	15
ISSN	1664-302X
DOIs	https://doi.org/10.3389/fmicb.2023.1121857
Publication status	Published - 2023

Keywords

calcium
characterization
cold adaptation
intracellular subtilisin protease
maturation
Planococcus
protein chemistry

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10.3389/fmicb.2023.1121857Licence: CC BY

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Rasmussen, C. B., Scavenius, C., Thøgersen, I. B., Harwood, S. L., Larsen, Ø., Bjerga, G. E. K., Stougaard, P., Enghild, J. J., & Thøgersen, M. S. (2023). Characterization of a novel cold-adapted intracellular serine protease from the extremophile Planococcus halocryophilus Or1. Frontiers in Microbiology, 14, Article 1121857. https://doi.org/10.3389/fmicb.2023.1121857

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title = "Characterization of a novel cold-adapted intracellular serine protease from the extremophile Planococcus halocryophilus Or1",

abstract = "The enzymes of microorganisms that live in cold environments must be able to function at ambient temperatures. Cold-adapted enzymes generally have less ordered structures that convey a higher catalytic rate, but at the cost of lower thermodynamic stability. In this study, we characterized P355, a novel intracellular subtilisin protease (ISP) derived from the genome of Planococcus halocryophilus Or1, which is a bacterium metabolically active down to -25°C. P355's stability and activity at varying pH values, temperatures, and salt concentrations, as well as its temperature-dependent kinetics, were determined and compared to an uncharacterized thermophilic ISP (T0099) from Parageobacillus thermoglucosidasius, a previously characterized ISP (T0034) from Planococcus sp. AW02J18, and Subtilisin Carlsberg (SC). The results showed that P355 was the most heat-labile of these enzymes, closely followed by T0034. P355 and T0034 exhibited catalytic constants (k cat ) that were much higher than those of T0099 and SC. Thus, both P355 and T0034 demonstrate the characteristics of the stability-activity trade-off that has been widely observed in cold-adapted proteases.",

keywords = "calcium, characterization, cold adaptation, intracellular subtilisin protease, maturation, Planococcus, protein chemistry",

author = "Rasmussen, {Casper B{\o}jer} and Carsten Scavenius and Th{\o}gersen, {Ida B} and Harwood, {Seandean Lykke} and {\O}ivind Larsen and Bjerga, {Gro Elin Kjaereng} and Peter Stougaard and Enghild, {Jan J} and Th{\o}gersen, {Mariane Schmidt}",

year = "2023",

doi = "10.3389/fmicb.2023.1121857",

language = "English",

volume = "14",

journal = "Frontiers in Microbiology",

issn = "1664-302X",

publisher = "Frontiers Media S.A",

}

Characterization of a novel cold-adapted intracellular serine protease from the extremophile Planococcus halocryophilus Or1. / Rasmussen, Casper Bøjer ; Scavenius, Carsten ; Thøgersen, Ida B et al.
In: Frontiers in Microbiology, Vol. 14, 1121857, 2023.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - Characterization of a novel cold-adapted intracellular serine protease from the extremophile Planococcus halocryophilus Or1

AU - Rasmussen, Casper Bøjer

AU - Scavenius, Carsten

AU - Thøgersen, Ida B

AU - Harwood, Seandean Lykke

AU - Larsen, Øivind

AU - Bjerga, Gro Elin Kjaereng

AU - Stougaard, Peter

AU - Enghild, Jan J

AU - Thøgersen, Mariane Schmidt

PY - 2023

Y1 - 2023

N2 - The enzymes of microorganisms that live in cold environments must be able to function at ambient temperatures. Cold-adapted enzymes generally have less ordered structures that convey a higher catalytic rate, but at the cost of lower thermodynamic stability. In this study, we characterized P355, a novel intracellular subtilisin protease (ISP) derived from the genome of Planococcus halocryophilus Or1, which is a bacterium metabolically active down to -25°C. P355's stability and activity at varying pH values, temperatures, and salt concentrations, as well as its temperature-dependent kinetics, were determined and compared to an uncharacterized thermophilic ISP (T0099) from Parageobacillus thermoglucosidasius, a previously characterized ISP (T0034) from Planococcus sp. AW02J18, and Subtilisin Carlsberg (SC). The results showed that P355 was the most heat-labile of these enzymes, closely followed by T0034. P355 and T0034 exhibited catalytic constants (k cat ) that were much higher than those of T0099 and SC. Thus, both P355 and T0034 demonstrate the characteristics of the stability-activity trade-off that has been widely observed in cold-adapted proteases.

AB - The enzymes of microorganisms that live in cold environments must be able to function at ambient temperatures. Cold-adapted enzymes generally have less ordered structures that convey a higher catalytic rate, but at the cost of lower thermodynamic stability. In this study, we characterized P355, a novel intracellular subtilisin protease (ISP) derived from the genome of Planococcus halocryophilus Or1, which is a bacterium metabolically active down to -25°C. P355's stability and activity at varying pH values, temperatures, and salt concentrations, as well as its temperature-dependent kinetics, were determined and compared to an uncharacterized thermophilic ISP (T0099) from Parageobacillus thermoglucosidasius, a previously characterized ISP (T0034) from Planococcus sp. AW02J18, and Subtilisin Carlsberg (SC). The results showed that P355 was the most heat-labile of these enzymes, closely followed by T0034. P355 and T0034 exhibited catalytic constants (k cat ) that were much higher than those of T0099 and SC. Thus, both P355 and T0034 demonstrate the characteristics of the stability-activity trade-off that has been widely observed in cold-adapted proteases.

KW - calcium

KW - characterization

KW - cold adaptation

KW - intracellular subtilisin protease

KW - maturation

KW - Planococcus

KW - protein chemistry

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U2 - 10.3389/fmicb.2023.1121857

DO - 10.3389/fmicb.2023.1121857

M3 - Journal article

C2 - 36910232

SN - 1664-302X

VL - 14

JO - Frontiers in Microbiology

JF - Frontiers in Microbiology

M1 - 1121857

ER -

Characterization of a novel cold-adapted intracellular serine protease from the extremophile Planococcus halocryophilus Or1

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Keywords

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