Extracellular proteases

Siddiqui lA, D Haas, S Heb (2005) Extracellular protease of Pseudomonas fluorescens CHAO, a biocontrol factor with activity against the root-knot nematode Meloidogyne incognita. Appl Environ Microbiol 71 5646-5649. 

The matrix metalloendoproteinases (MMPs or matrixins) are a family of zinc and calcium dependent extracellular proteases that collectively degrade most of the protein constituents of the extracellular matrix [9]. There are at least 23 members of this family and are divided primarily on the basis of sequence homology and substrate specificity into the following grouping collagenases (MMP-1, -8, -13, -18) gelatinases... 

B. brevis is not so well studied. It also shows low extracellular protease activity, and a protease-deficient strain is available [43]. High [44] and low [45] copy number plasmids are constructed for different levels of expression. Modification of signal sequences can enhance yields of eukaryotic proteins [46]. The yields can be up to 3 gL-1 media. 

Staphylococcus carnosus is genetically highly stable, a good secretor (Gram positive) able to translocate proteins containing several hydrophobic transmembrane regions [58], and it has no extracellular proteases, which makes it suitable for production of the secreted enzymes [58],... 

Wu, X.C., Lee, W., Tran, L. and Wong, S.L. (1991) Engineering a Bacillus subtilis expression-secretion system with a strain deficient in six extracellular proteases. Journal of Bacteriology, 173 (16), 4952 1958. 

Microorganisms may be capable of metabolizing the product itself, thus reducing its potency. This is particularly true of protein-based biopharmaceuticals, as most microbes produce an array of extracellular proteases. 

Regulation of neuropeptide expression is exerted at several levels. Control of neuropeptide function is mediated by factors controlling rates of prepropeptide gene transcription, translation, peptide degradation and secretion (Fig. 18-11). On the scale of seconds to minutes, peptide secretion is not always coupled lock-step with classical transmitter release (example above). Peptides are inactivated by diffusion and by proteolysis, so it would be expected that inhibition of specific extracellular proteases... 

H 0 A-butanoyl-L-homoserine lactone, BHL or C4-HSL Aeromomas hydrophila Aeromonas salmonicida Pseudomonas aeruginosa, Serratia liquefaciens Extracellular protease, biofilm formation. Extracellular protease. Virulence factors - alkaline protease, cyanide, elastase, haemolysin, lectins, pyocyanin, rhaminolipid, RpoS Swarming, protease. 

Gomez-Duran A, Mulero-Navarro S, Chang X, Fernandez-Salguero PM LTBP-1 blockade in dioxin receptor-null mouse embryo fibroblasts decreases TGF-(5 activity role of extracellular proteases plasmin and elastase. J Cell Biochem 2006 97 380-392. 

In fact, halophilic enzymes have been shown to function in low salt environments, where the salt is replaced by a suitable organic solvent typically those that are kosmotropic. For example, an extracellular protease from H. halobium, which requires 4M NaCl for optimal function, is highly active in as little as 0.2 M NaCl in the presence of 40% (v/v) dimethylsulfoxide (DMSO) [10]. The behavior of salts... 

Figure 3.2 Replacement of high salt with organic solvents-stability of the extracellular protease from Halobacterium halobium in 0.36 M NaCl in water or various organic solvents. The KSC value represents the Sechenow constant multiplied by the concentration of the solvent, which provides a measure of the salting-out (kosmotropic) capacity of the solution [10].

Tanaka, T. Kawata, M. Gloning and characterization of Bacillus subtilis iep, which has positive and negative effects on production of extracellular proteases. J. Bacteriol., 170, 3593-3600 (1988)... 

Psychrotrophic Pseudomonas spp. are capable of producing extracellular proteases, glycosidases, and phospholipases which can degrade various components of the milk fat globule membrane, making it more susceptible to lipolysis (Alkanhal et al 1985 Chrisope and Marshall 1976 Marin et al. 1984). 

Alichanidis, E. and Andrews, A. T. 1977. Some properties of the extracellular protease produced by the psychrotrophic bacterium Pseudomonas fluorescens strain AR-11. Biochim. Biophys. Acta 485, 424-433. 

Juan, S. M. and Cazzulo, J. J. 1976. The extracellular protease from Pseudomonas fluore-scens. Experientia 32, 1120-1122. 

Modler, H., Brunner, J. R. and Stine, C. M. 1974. Extracellular protease of Pencillium roqueforti. II. Characterization of a purified enzyme preparation. J. Dairy Sci. 57, 528-534. 

The first pyrolysin to be cloned from a higher plant was cucumisin from Cucumis melo, an extracellular protease highly abundant in melon fruit [152], Cucumisin was shown to have a broad substrate specificity in that it cleaves a variety of small peptide substrates and eight peptide bonds within the oxidized insulin B chain [153-155]. A similar, broad... 

In chapter 8, we mentioned that the pancreas secretes trypsin, chymotrypsin, and elastase as inactive zymogens, which are activated by extracellular proteases. Trypsin is activated when the intestinal enzyme enter-opeptidase cuts off an N-terminal hexapeptide. Trypsin in turn activates chymotrypsin by cutting it at the N-terminal end between Arg 15 and lie 16. This type of change in the covalent structure of an enzyme is termed partial proteoly-sis. Delaying the activation prevents the digestive enzymes from destroying the pancreatic cells in which they are synthesized. 

Extracellular proteases are of commercial value and find multiple applications in various industrial sectors. A good number of bacterial alkaline proteases are commercially available, such as Subtilisin Carlsberg, subtilisin BPN and Savinase, with their major application as detergent enzymes. 

The aim of this chapter is to summarize the information available about the extracellular proteases of entomopathogenic fungi, focusing on Metarhizium and Beauveria species. A large number of protease enzymes have been purified, cloned and characterized from these beneficial organisms. These results will be reviewed and data about the regulation of proteases as well as their role during the biocontrol process will be discussed. 

Several extracellular proteases have been purified and characterized from entomopathogenic fungi. Molecular weights, isoelectric points and specific inhibitors of some of these enzymes are summarized in Table 1. The majority of them belong to the family of serine proteases. 

Table 1. Properties of extracellular proteases purified from entomopathogenic fungi... 

Cloning of Extracellular Protease Encoding Genes from... 

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