Purification and Characterization of a Cold Alkaline Protease from a Psychrophilic Pseudomonas aeruginosa HY1215

Hao, Jianhua; Sun, Ming

doi:10.1007/s12010-014-1315-2

Cited by 16 publications

(17 citation statements)

References 22 publications

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“…A few species of bacteria have several metalloprotease genes in their genomes [12], [22], although the reason for multiple serralysin family metalloproteases is unclear, given that these proteins generally have broad substrate specificity and maintain functionality over a reasonably broad range of temperature and pH [15], [33]. There are four predicted metalloprotease genes in the S. marcescens reference strain Db11 genome [27].…”

Section: Introductionmentioning

confidence: 99%

SlpE is a calcium-dependent cytotoxic metalloprotease associated with clinical isolates of Serratia marcescens

Stella

Callaghan

Zhang

et al. 2017

Research in Microbiology

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Serralysin-like proteases are found in a wide variety of bacteria. These metalloproteases are frequently implicated in virulence and are members of the widely conserved RTX-toxin family. We identified a serralysin-like protease in the genome of a clinical isolate of S. marcescens that is highly similar to the canonical serralysin protein, PrtS. This gene was named serralysin-like protease E, SlpE, and was found in the majority (67%) of tested clinical isolates, but was absent from most tested non-clinical isolates including the insect pathogen and reference S. marcescens strain Db11. Purified recombinant SlpE exhibited calcium-dependent protease activity similar to metalloproteases PrtS and SlpB. Induction of slpE in the low-protease-producing S. marcescens strain PIC3611 highly elevated extracellular protease activity, and extracellular secretion required the lipD type 1 secretion system gene. Transcription of slpE was highly reduced in an eepR transcription factor mutant. Mutation of the slpE gene in a highly proteolytic clinical isolate reduced its protease activity, and evidence suggests that SlpE confers cytotoxicity of S. marcescens to the A549 airway carcinoma cell line. Together, these data reveal SlpE to be an EepR-regulated cytotoxic metalloprotease associated with clinical isolates of an important opportunistic pathogen.

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Section: Introductionmentioning

confidence: 99%

SlpE is a calcium-dependent cytotoxic metalloprotease associated with clinical isolates of Serratia marcescens

Stella

Callaghan

Zhang

et al. 2017

Research in Microbiology

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“…One hundred microliters of enzyme solution were mixed with 4.9 mL of casein solution (0.6% ( w / v ) in 25 mM borate buffer, pH 10.0) and incubated at 25 °C for 10 min. The relative enzyme activity was measured using Folin-Ciocalteu’s method [ 3 , 4 ]. One unit was defined as the amount of enzyme causing the release of 1 μg tyrosine per minute under the above conditions.…”

Section: Methodsmentioning

confidence: 99%

“…Proteases are the most important industrial enzymes, accounting for more than 60% of the total enzyme market [ 2 , 3 ]. They have broad applications in the pharmaceutical, leather, food, and detergent industries [ 1 , 2 , 3 , 4 ]. Proteases play critical roles in normal biological processes; their unusual activities have been implicated in the development and progression of many diseases, e.g., fibrosis, arthritis, cancer, cardiovascular diseases, nephritis, and central nervous system disorders [ 5 , 6 , 7 ].…”

Section: Introductionmentioning

confidence: 99%

Structure-Based Design and Synthesis of a New Phenylboronic-Modified Affinity Medium for Metalloprotease Purification

Wang²,

Xu³

et al. 2016

Marine Drugs

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Metalloproteases are emerging as useful agents in the treatment of many diseases including arthritis, cancer, cardiovascular diseases, and fibrosis. Studies that could shed light on the metalloprotease pharmaceutical applications require the pure enzyme. Here, we reported the structure-based design and synthesis of the affinity medium for the efficient purification of metalloprotease using the 4-aminophenylboronic acid (4-APBA) as affinity ligand, which was coupled with Sepharose 6B via cyanuric chloride as spacer. The molecular docking analysis showed that the boron atom was interacting with the hydroxyl group of Ser176 residue, whereas the hydroxyl group of the boronic moiety is oriented toward Leu175 and His177 residues. In addition to the covalent bond between the boron atom and hydroxyl group of Ser176, the spacer between boronic acid derivatives and medium beads contributes to the formation of an enzyme-medium complex. With this synthesized medium, we developed and optimized a one-step purification procedure and applied it for the affinity purification of metalloproteases from three commercial enzyme products. The native metalloproteases were purified to high homogeneity with more than 95% purity. The novel purification method developed in this work provides new opportunities for scientific, industrial and pharmaceutical projects.

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“…In recent times, Antarctic cold habitats such as deep-sea sediment, Prydz Bay [ 11 ]; soil samples, King George Island [ 12 – 15 ]; seawater and krill [ 16 ]; marine water, Casey Station [ 17 ]; and penguin feathers [ 18 ] have been explored for isolation of cold-adapted protease-producing microorganisms. Other cold environments inhabiting cold-adapted protease-producing microorganisms are Japanese deep-sea water [ 19 ], yellow sea [ 20 , 21 ], Ikka Fjord and surficial sediments in Greenland [ 22 ], deep-sea mud in Eastern Indian Ocean [ 23 ], and Bolu Mountain and Erzurum soil in Turkey [ 24 , 25 ]. Cold-adapted protease producers have been isolated from soil samples of various Indian Himalayan regions such as Lahaul and Spiti [ 26 ], Kashmir apple garden [ 27 ], Thajiwas glacier [ 28 ], Gangotri glacier [ 29 ], and Wular Lake [ 30 ].…”

Section: Main Textmentioning

confidence: 99%

Adaptation, production, and biotechnological potential of cold-adapted proteases from psychrophiles and psychrotrophs: recent overview

Furhan

2020

Journal of Genetic Engineering and Biotechnology

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Background Proteases or peptidases are an imperative class of hydrolytic enzymes capable of hydrolyzing large proteins into smaller peptides. The cold-adapted proteases show higher catalytic capacity in low temperatures as well as stability in alkaline conditions and appear as strong contenders for various applications in special industries. Main body In the past few decades, the interest in cold-adapted microorganisms producing cold-adapted proteases has increased at an exciting rate, and many of them have emerged as important biotechnological and industrial candidates. Industrial proteases are largely supplied from various types of microorganisms than plant or animal sources. Among diverse microbial sources, psychrophiles and psychrotrophs inhabiting permanently or partially cold environments have appeared as rich sources of cold-adapted proteases. Short conclusion The present review focuses on recent sources of cold-adapted protease producers along with the molecular adaptation of psychrotrophs and psychrophiles. The recent knowledge on production, kinetic properties, purification, and substrate specificity of cold-adapted proteases has been summarized. Recent advances in cold-adapted protease gene cloning and structural studies are also described. Moreover, the prospective applications of cold-adapted proteases are discussed which can help in evaluating their industrial potential.

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Purification and Characterization of a Cold Alkaline Protease from a Psychrophilic Pseudomonas aeruginosa HY1215

Cited by 16 publications

References 22 publications

SlpE is a calcium-dependent cytotoxic metalloprotease associated with clinical isolates of Serratia marcescens

SlpE is a calcium-dependent cytotoxic metalloprotease associated with clinical isolates of Serratia marcescens

Structure-Based Design and Synthesis of a New Phenylboronic-Modified Affinity Medium for Metalloprotease Purification

Adaptation, production, and biotechnological potential of cold-adapted proteases from psychrophiles and psychrotrophs: recent overview

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