A novel halolysin without C-terminal extension from an extremely halophilic archaeon

Hou, Jing; Li, Si-Ya; Zhao, Yang-Jie; Cui, Heng‐Lin

doi:10.1007/s00253-022-11903-4

Cited by 7 publications

(4 citation statements)

References 31 publications

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“…magadii was >30% of original activity after incubation at 60°C for 2 h (Gimeńez et al, 2000), that of Hly Hap from Haladaptatus sp. DYF46 was 33.9% after incubation at 60°C for 1 h (Hou et al, 2022), and that of Hly from Hgn. rubrum RO2-11 was <20% after incubation at 60°C for 1 h (Gao et al, 2017).…”

Section: Effect Of Temperature Ph and Nacl Concentration On Hslhlyb A...mentioning

confidence: 92%

“…J7 (Zhang et al, 2014;Du et al, 2015), Nep from Natrialba magadii (De Castro et al, 2008), HlyA from Halococcus salifodinae (Hou et al, 2020), Hly Hap from Haladaptatus sp. DYF46 (Hou et al, 2022). For protein domain organization, those halolysin precursors are usually made up of a Tat (twin-arginine translocation) signal peptide, an N-terminal propeptide, an S8/S53 family catalytic domain and a C-terminal extension (CTE).…”

Section: Open Access Edited Bymentioning

confidence: 99%

“…The CTE of halolysins are usually prepeptidase C-terminal (PPC) domain, except a combined polycystic kidney disease (PKD) domain and chitin-binding domain (ChBD) was reported in the CTE of SptC (Du et al, 2015). In recent years a marine halolysin Hly Hap without CTE was discovered to prefer a low salinity in contrast to most halolysins (Hou et al, 2022). CTE domains have been shown to have a significant impact on the structure and function of a protease (Gao et al, 2010), especially those secreted by the marine bacteria (Huang et al, 2017).…”

Section: Open Access Edited Bymentioning

confidence: 99%

“…Culture supernatant (12,000 g, 10 min, 4°C) was used as the crude enzyme to assay the extracellular protease activity. To obtain purified enzymes, the HslHlyB and HslHlyBDCTE precursors were expressed in E. coli BL21 (DE3) and in vitro refolded as previously described (Hou et al, 2021;Hou et al, 2022). For in vitro refolding, the recombinant cells were firstly resuspended in lysis buffer (8 M urea, 10 mM CaCl 2 , 50 mM Tris-HCl, 2 mM b-mercaptoethanol, pH 8.0).…”

Section: Expression and Purification Of Halolysinsmentioning

confidence: 99%

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An extracellular protease containing a novel C-terminal extension produced by a marine-originated haloarchaeon

2023

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Marine microorganisms have long been acknowledged as a significant reservoir of enzymes required for industrial use. In this study, a novel extracellular protease HslHlyB derived from marine-originated haloarchaeon Halostella pelagica DL-M4T was identified. HslHlyB contained polycystic kidney disease (PKD) domain and pre-peptidase C-terminal (PPC) domain at the C-terminus. Truncation and replacement of the C-terminal extension (CTE) of HslHlyB demonstrated the importance of the CTE in maintaining the protease activity secreted by haloarchaeon. HslHlyB and HslHlyBΔCTE were expressed in Escherichia coli BL21(DE3), and purified by high-affinity column refolding and gel filtration chromatography. The molecular masses of HslHlyB and HslHlyBΔCTE were 42 kDa and 20 kDa, respectively. The optimum catalytic reaction conditions were 50°C, pH 8.5, NaCl 3.5 M and 50°C, pH 7.5, NaCl 3 M, respectively. They showed good stability and hydrolysis capabilities towards a wide range of protein substrates. HslHlyBΔCTE showed higher catalytic reaction rate and better thermal stability than the wild type against azocasein and tetrapeptide substrate. The hydrolysates of soybean protein hydrolyzed by HslHlyBΔCTE had smaller average molecular masses and shorter average peptide chain lengths than those by HslHlyB. These results indicated the diversity of halolysins from marine-originated haloarchaea to harness organic nitrogen in the marine environment and provided promising candidates for application in various industries.

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Section: Effect Of Temperature Ph and Nacl Concentration On Hslhlyb A...mentioning

confidence: 92%

Section: Open Access Edited Bymentioning

confidence: 99%

Section: Open Access Edited Bymentioning

confidence: 99%

Section: Expression and Purification Of Halolysinsmentioning

confidence: 99%

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An extracellular protease containing a novel C-terminal extension produced by a marine-originated haloarchaeon

2023

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Extracellular proteases from halophiles: diversity and application challenges

Nwankwo

Hou

Cui

2023

Appl Microbiol Biotechnol

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Hly176B, a low-salt tolerant halolysin from the haloarchaeon Haloarchaeobius sp. FL176

Zhang

Chen

et al. 2023

World J Microbiol Biotechnol

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Extracellular proteases of haloarchaea are adapted to function at high concentrations of NaCl, and could nd useful applications in industrial or biotechnological processes where hypersaline conditions are desired. The diversity of extracellular proteases produced by haloarchaea is largely unknown, although the genomes of many species have been sequenced and are publicly available. In this study, a gene encoding the extracellular protease Hly176B from the haloarchaeon Haloarchaeobius sp. FL176 was cloned and expressed in Escherichia coli. The catalytic triad Asp-His-Ser was con rmed via site-directed mutagenesis, indicating that Hly176B belongs to the class of serine proteases (halolysin). Unlike previously reported extracellular proteases from haloarchaea, Hly176B remained active in almost salt-free solution for a relatively long time. In addition, Hly176B displayed good tolerance to some metal ions, surfactants and organic solvents, and exerts its highest enzyme activities at 40 °C, pH 8.0 and 0.5 M NaCl. Therefore, this study enriches our knowledge of extracellular proteases, and also expands their applications in various industrial uses.

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A novel halolysin without C-terminal extension from an extremely halophilic archaeon

Cited by 7 publications

References 31 publications

An extracellular protease containing a novel C-terminal extension produced by a marine-originated haloarchaeon

An extracellular protease containing a novel C-terminal extension produced by a marine-originated haloarchaeon

Extracellular proteases from halophiles: diversity and application challenges

Hly176B, a low-salt tolerant halolysin from the haloarchaeon Haloarchaeobius sp. FL176

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