Isolation, purification and characterization of a pH tolerant and temperature stable proteinaceous protease inhibitor from marine Pseudomonas mendocina

Sapna, K.; Ali, Pervaiz; Mol, K. R. Rekha; Bhat, Sarita G.; Chandrasekaran, M.; Elyas, K. K.

doi:10.1007/s10529-017-2424-0

Cited by 4 publications

(4 citation statements)

References 17 publications

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“…The plate was incubated at 37°C. The inhibitory activity was detected by absence of clearance zone around the well containing the inhibitor (Sapna, 2013).…”

Section: Primary Screening Of Protease Inhibitory Activity Of Proteinmentioning

confidence: 99%

Expression of a novel gene encoding protease inhibitor from metagenome of sponge in Vietnam

Hong¹,

Dat²,

Hoa³

et al. 2019

V J Bio

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Marine sponge is known as a “gold mine” of natural products from marine environment. Many novel bioactive compounds have been isolated from marine sponges and sponge-associated microorganisms such as antibiotics, anti-cancer compounds, protease inhibitors, etc. In this study, we selected a gene encoding protease inhibitor from metagenome of a sponge collected in Da Nang to express in Escherichia coli BL21(DE3). The gene PI-DN9 encoding protease inhibitor (1.3 kb) was cut off cloning vector pUC57/PI-DN9 containing gene PI-DN9 and inserted into expression vector pET32a(+), the recombinant vector pET32a(+)/PI-DN9 then was transformed and expressed in the E. coli strain BL21(DE3). Results showed that recombinant protein (50 kDa) was expressed successfully at 25°C, 1 mM of IPTG in 5 hours. The recombinant protein was purified using Ni-NTA affinity chromatography column. Western blot assay and bioactive assay showed good activity of the purified protein.

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“…The plate was incubated at 37°C. The inhibitory activity was detected by absence of clearance zone around the well containing the inhibitor (Sapna, 2013).…”

Section: Primary Screening Of Protease Inhibitory Activity Of Proteinmentioning

confidence: 99%

Expression of a novel gene encoding protease inhibitor from metagenome of sponge in Vietnam

Hong¹,

Dat²,

Hoa³

et al. 2019

V J Bio

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show abstract

“…Proteinaceous inhibitors are ubiquitous inhibitors and isolated from different sources (e.g., microorganisms, plants, and animals). Natural proteinaceous inhibitors are known as templates for the modification of natural control mechanisms and as a source of basic design principles [29]. Proteinaceous inhibitors are usually classified based on the kind of inhibited protease.…”

Section: Proteinaceous Inhibitorsmentioning

confidence: 99%

“…The metal ions play an important role in maintaining the structure of PIs. The structural stability of proteins is enhanced by divalent metal ions as the metal ions can attain the critical conformation that is needed for biological activity of the protein [29]. For example, cysteine protease inhibitor from pearl millet needs the Zn 2+ for the protease inhibitory and antifungal activity of the protein [37].…”

Section: Effect Of Metal Ionsmentioning

confidence: 99%

“…Some detergents (e.g., cationic, anionic, zwitterionic and non-ionic) are used for solubilizing proteins from lipid membranes. Therefore, proteinase inhibitors are often combined with detergents in cell lysis buffers to inhibit undesired proteolysis and facilitate membrane protein solubilization in protein purification procedures [29]. Normally, nonionic detergents are considered as mild detergents and not interact extensively with the protein surface, while ionic detergents (e.g., SDS) generally bind unwanted to the protein surface, which results in protein unfolding [46].…”

Section: Effect Of Detergentsmentioning

confidence: 99%

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Mini-Review PROTEASE INHIBITOR (PI) AND PIs FROM SPONGE-ASSOCIATED MICROORGANISMS

Hong¹,

Dat²,

Cúc³

et al. 2018

JST

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Protease inhibitors (PIs) are proteins or peptides inhibiting the activity of protease and constitute a very important mechanism for regulating protease activity. So far, PIs have been used for the study of enzyme structures and reaction mechanisms, but recently they have been used in pharmaceutical, agricultural and industrial fields. Compared to terrestrial counterparts, marine environment possesses their unique characters, therefore, they are capable of producing a wide range of novel bioactive compounds including protease inhibitors. In our review, a brief overview of protease inhibitors (e.g., classification, mechanisms, and characteristics of PIs) and protease inhibitors from marine sponges and sponge-associated microorganisms has been reviewed.

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Potential Anticoagulant Activity of Trypsin Inhibitor Purified from an Isolated Marine Bacterium Oceanimonas Sp. BPMS22 and its Kinetics

Harish

Uppuluri

2018

Mar Biotechnol

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Protease inhibitors control major biological protease activities to maintain physiological homeostasis. Marine bacteria isolated from oligotrophic conditions could be taxonomically distinct, metabolically unique, and offers a wide variety of biochemicals. In the present investigation, marine sediments were screened for the potential bacteria that can produce trypsin inhibitors. A moderate halotolerant novel marine bacterial strain of Oceanimonas sp. BPMS22 was isolated, identified, and characterized. The effect of various process parameters like salt concentration, temperature, and pH was studied on the growth of the bacteria and production of trypsin inhibitor. Further, the trypsin inhibitor was purified to near homogeneity using anion exchange, size exclusion, and affinity chromatography. The purified trypsin inhibitor was found to competitively inhibit trypsin activity with an inhibition coefficient, K, of 3.44 ± 0.13 μM and second-order association rate constant, k, of 1.08 × 10 M S. The proteinaceous trypsin inhibitor had a molecular weight of approximately 30 kDa. The purified trypsin inhibitor showed anticoagulant activity on the human blood samples.

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Isolation, purification and characterization of a pH tolerant and temperature stable proteinaceous protease inhibitor from marine Pseudomonas mendocina

Abstract: A pH tolerant and thermostable protease inhibitor BTPI-301 active against trypsin was purified and characterized from P. mendocina that could be developed and used as biopreservative as well as biocontrol agent.

Cited by 4 publications

References 17 publications

Expression of a novel gene encoding protease inhibitor from metagenome of sponge in Vietnam

Expression of a novel gene encoding protease inhibitor from metagenome of sponge in Vietnam

Mini-Review PROTEASE INHIBITOR (PI) AND PIs FROM SPONGE-ASSOCIATED MICROORGANISMS

Potential Anticoagulant Activity of Trypsin Inhibitor Purified from an Isolated Marine Bacterium Oceanimonas Sp. BPMS22 and its Kinetics

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