Microbial proteases: Production and application in obtaining protein hydrolysates

Aguilar, Jessika Gonçalves dos Santos; Sato, Hélia Harumi

doi:10.1016/j.foodres.2017.10.044

Cited by 167 publications

(66 citation statements)

References 132 publications

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“…Bacillus proteases have several remarkable characteristics for many industrial applications such as broad pH and temperature activity, stability range, alkaline and toxic compounds tolerance, including oxidants and surfactants ( Contesini et al, 2018 ). Based on these interesting aspects, proteases from the genus Bacillus are used in the production of detergents, food products, leather, silk, and agrochemical products ( Dos Santos Aguilar and Sato, 2018 ; Razzaq et al, 2019 ). The alkaline serine protease (DHAP) from Bacillus pumilus BA06 is a typical mesophilic enzyme, which has demonstrated great potential in various industrial applications ( Zhao and Feng, 2018 ).…”

Section: Introductionmentioning

confidence: 99%

The Practical Potential of Bacilli and Their Enzymes for Industrial Production

Danilova

Шарипова

2020

Front. Microbiol.

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Bacillus spp. are an affordable source of enzymes due to their wide distribution, safety in work, ease of cultivation, and susceptibility to genetic transformations. Researchers are particularly interested in proteolytic enzymes, which constitute one of the most diverse groups of microbial proteins in terms of properties. Despite the long history of their research, this group of enzymes continue to show great potential for practical application in the biomedical industry, as well as in the agricultural industry. Thus, the unique properties of bacillary proteinases, such as stability in a wide range of temperatures and pH, high specificity, biodegradability of a wide range of substrates, and the high potential of sequenced Bacillus genomes are a powerful foundation for the development of new biotechnologies. The current review aims to discuss recent studies on various enzymes in particular, proteinases produced by bacteria of the genus Bacillus , along with their prospective practical applications. This article also presents an interpretive summary of the recent developments on the usage of probiotic Bacillus strains as potential feed additives.

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Section: Introductionmentioning

confidence: 99%

The Practical Potential of Bacilli and Their Enzymes for Industrial Production

Danilova

Шарипова

2020

Front. Microbiol.

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“…The culture of protease-producing microorganisms in high quantities and in a short time can be done using established fermentation methods. Furthermore, microorganisms facilitate the production of proteases with broad natural biochemical diversity and engineering of novel enzymes through genetic manipulations [3,4].…”

Section: Introductionmentioning

confidence: 99%

Structural and biochemical characterization of a novel thermophilic Coh01147 protease

et al. 2020

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Proteases play an essential role in living organisms and represent one of the largest groups of industrial enzymes. The aim of this work was recombinant production and characterization of a newly identified thermostable protease 1147 from thermophilum indigenous Cohnella sp. A01. Phylogenetic tree analysis showed that protease 1147 is closely related to the cysteine proteases from DJ-1/ThiJ/PfpI superfamily, with the conserved catalytic tetrad. Structural prediction using MODELLER 9v7 indicated that protease 1147 has an overall α/β sandwich tertiary structure. The gene of protease 1147 was cloned and expressed in Escherichia coli (E. coli) BL21. The recombinant protease 1147 appeared as a homogenous band of 18 kDa in SDS-PAGE, which was verified by western blot and zymography. The recombinant protein was purified with a yield of approximately 88% in a single step using Ni-NTA affinity chromatography. Furthermore, a rapid one-step thermal shock procedure was successfully implemented to purify the protein with a yield of 73%. Using casein as the substrate, K m , and k cat , k ca t/K m values of 13.72 mM, 3.143 × 10 −3 (s-1), and 0.381 (M-1 S-1) were obtained, respectively. The maximum protease activity was detected at pH = 7 and 60˚C with the inactivation rate constant (kin) of 2.10 × 10-3 (m-1), and half-life (t 1/2) of 330.07 min. Protease 1147 exhibited excellent stability to organic solvent, metal ions, and 1% SDS. The protease activity was significantly enhanced by Tween 20 and Tween 80 and suppressed by cysteine protease specific inhibitors. Docking results and molecular dynamics (MD) simulation revealed that Tween 20 interacted with protease 1147 via hydrogen bonds and made the structure more stable. CD and fluorescence spectra indicated structural changes taking place at 100˚C, very basic and acidic pH, and in the presence of Tween 20. These properties make this newly characterized protease a potential candidate for various biotechnological applications.

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“…In contrast, other proteases demonstrate high specificity, often related to the tight control of a cellular or physiological process (4). Because of their unique activity, proteases have a wide range of applications in biotechnology and the food industry (5)(6)(7).…”

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confidence: 99%

Discovery of a new Pro-Pro endopeptidase, PPEP-2, provides mechanistic insights into the differences in substrate specificity within the PPEP family

Klychnikov

Shamorkina

Weeks

et al. 2018

Journal of Biological Chemistry

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Pro-Pro endopeptidases (PPEPs) belong to a recently discovered family of proteases capable of hydrolyzing a Pro-Pro bond. The first member from the bacterial pathogen (PPEP-1) cleaves two cell-surface proteins involved in adhesion, one of which is encoded by the gene adjacent to the gene. However, related PPEPs may exist in other bacteria and may shed light on substrate specificity in this enzyme family. Here, we report on the homolog of PPEP-1 in, which we denoted PPEP-2. We found that PPEP-2 is a secreted metalloprotease, which likewise cleaved a cell-surface protein encoded by an adjacent gene. However, the cleavage motif of PPEP-2, PLP↓PVP, is distinct from that of PPEP-1 (VNP↓PVP). As a result, an optimal substrate peptide for PPEP-2 was not cleaved by PPEP-1 and vice versa. To gain insight into the specificity mechanism of PPEP-2, we determined its crystal structure at 1.75 Å resolution and further confirmed the structure in solution using small-angle X-ray scattering (SAXS). We show that a four-amino-acid loop, which is distinct in PPEP-1 and -2 (GGST in PPEP-1 and SERV in PPEP-2), plays a crucial role in substrate specificity. A PPEP-2 variant, in which the four loop residues had been swapped for those from PPEP-1, displayed a shift in substrate specificity toward PPEP-1 substrates. Our results provide detailed insights into the PPEP-2 structure and the structural determinants of substrate specificity in this new family of PPEP proteases.

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Microbial proteases: Production and application in obtaining protein hydrolysates

Cited by 167 publications

References 132 publications

The Practical Potential of Bacilli and Their Enzymes for Industrial Production

The Practical Potential of Bacilli and Their Enzymes for Industrial Production

Structural and biochemical characterization of a novel thermophilic Coh01147 protease

Discovery of a new Pro-Pro endopeptidase, PPEP-2, provides mechanistic insights into the differences in substrate specificity within the PPEP family

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