Trypsin alkaline phosphatase

Several enzymes have been immobilized in sol-gel matrices effectively and employed in diverse applications. Urease, catalase, and adenylic acid deaminase were first encapsulated in sol-gel matrices [72], The encapsulated urease and catalase retained partial activity but adenylic acid deaminase completely lost its activity. After three decades considerable attention has been paid again towards the bioencapsulation using sol-gel glasses. Braun et al. [73] successfully encapsulated alkaline phosphatase in silica gel, which retained its activity up to 2 months (30% of initial) with improved thermal stability. Further Shtelzer et al. [58] sequestered trypsin within a binary sol-gel-derived composite using TEOS and PEG. Ellerby et al. [74] entrapped other proteins such as cytochrome c and Mb in TEOS sol-gel. Later several proteins such as Mb [8], hemoglobin (Hb) [56], cyt c [55, 75], bacteriorhodopsin (bR) [76], lactate oxidase [77], alkaline phosphatase (AP) [78], GOD [51], HRP [79], urease [80], superoxide dismutase [8], tyrosinase [81], acetylcholinesterase [82], etc. have been immobilized into different sol-gel matrices. Hitherto some reports have described the various aspects of sol-gel entrapped biomolecules such as conformation [50, 60], dynamics [12, 83], accessibility [46], reaction kinetics [50, 54], activity [7, 84], and stability [1, 80],... 

Kai and Imakubo(76) found that the temperature at which emission from the exposed tryptophan is no longer observed appears to be characteristic of the protein, having values of 180 K for trypsin, 200 K for aldolase, and 230 K for alkaline phosphatase. 

Inhibits serine proteases such as trypsin and chymotrypsin. Also inhibits cysteine proteases (reversible by reduced thiols) and mammalian acetylcholinesterase Inhibits ATPase, alkaline phosphatase and tyrosine phosphatase Reagent for maintaining -SH groups in the reduced state. Effective for reducing protein disulfide bonds prior to SDS-PAGE... 

Minhas T and Greenman J (1989) Production of cell-bound and vesicle-associated trypsin-like protease, alkaline phosphatase and N-acetyl—glucosaminidase by Bacteroides gingivalis strain W50. J Gen Microbiol 135, 557-564. 

FIGURE 2-21 The pH optima of some enzymes. Pepsin is a digestive enzyme secreted into gastric juice trypsin, a digestive enzyme that acts in the small intestine alkaline phosphatase of bone tissue, a hydrolytic enzyme thought to aid in bone mineralization. 

Rothman and Byrne 46) have used tryptic digestion to determine whether the subunits of alkaline phosphatase are identical. Since trypsin specifically cleaves at lysyl and arginyl residues, there will be as many peptides formed as there are arginine and lysine residues if the mono-... 

An alternative displacement method, reported by Nagao and coworkers (46,47), is based on a lengthy and more complex procedure. Sarin-bound acetylcholinesterase was solubilized from erythrocyte membranes, digested with trypsin (37 °C, 24 h), and the hydrolysis product isopropyl methylphos-phonic acid released by digestion with alkaline phosphatase (37 °C, 48 h). High molecular mass... 

The influence of the ovarian cycle on protease activity in the vagina has also been demonstrated. For example, the trypsin-like activity in rat vaginal smears was found to be maximal at proestrus. The activity of //-glucuronidase, acid phosphatase, alkaline phosphatase, and esterase all vary in the vaginal tissue of premenopausal and postmenopausal women. 

An alternate application of the SAS technology was to fractionally precipitate alkaline phosphatase, insulin, ribonuclease and trypsin mixtures from DMSO (Winters et al., 1999). Separation and purihcation of the proteins were achieved by their differing solubility in COg expanded DMSO. Substantial biological activity was recovered for insulin, lysozyme, ribonuclease and trypsin however alkaline phosphatase was irreversibly denatured. 

Winters et al. (87) studied the GAS precipitation of alkaline phosphatase, catalase, lysozyme, ribonuclease, and trypsin from neat DMSO using pressurized gaseous CO2 at 34°C as the antisolvent. The activity of the reconstituted proteins was measured and was found to decrease with increasing molecular weight (Fig. 4). Thus, although ribonuclease (13.7 kDa) regained... 

In the case of sarin-inhibited ChE, the phos-phyl moiety has also been displaced as isopropyl methylphosphonic acid using trypsin digestion and alkaline phosphatase (Nagao et al., 1997 Matsuda et al., 1998). 

Abbreviations are LY, hen egg-white lysozyme CON A, demetallized concanavalin A TP, demetallized porcine trypsin tRNA, nonspecific yeast transfer ribonucleic acid CA, human erythrocyte carbonic anhydrase B Hb, human adult carbonmonoxyhemo-globin AP, E. coli alkaline phosphatase TF, demetallized human transferrin IG, human nonspecific -/-immunoglobulin AD, alcohol dehydrogenase from yeast CP, human ceruloplasmin HC1 / 20, l/IO(L), 1/10(0, 1/2, 1/1, various states of association of Helix pomatia hemocyanin. Dashed line m>calculated using Equation 3 with no adjustable parameteis, using the viscosity of pure water to compute v . The proteins were assumed spherical, and a 3J-A hydration layer was included in computing the hydrodynamic radii. After Ref. 7. 

Note References used are Craik et al. (1987) for rat trypsin Carter and Wells (1988) for subtilisin Nickbarg et al. (1988) for yeast trios hosphate isomerase (GAP for glyceraldehyde-3-phosphate and DHAP for dihydroxyacetone phosphate) Soukri et al. (1989) for E. coli glyceraldehyde-3-phosphate dehydrogenase Kim and Patel (1992) for human lipoamide dehydrogenase Fan et al. (1991) for yeast alcohol dehydrogenase Scrutton et al. (1990) for E. coli glutathione reductase Murphy et al. (1993) for E. coli alkaline phosphatase Leatherbarrow et al. (1985) for tyrosyl-tRNA synthetase. 

Figure 5 Correlation between radius of entrapped enzyme and that of a microemulsion droplet at maximum activity. The following enzymes were investigated 1, lysozyme 2, cytochrome c 3, trypsin 4, a-chymotrypsin 5, ribonuclease 6, pepsin 7, lipase 8, peroxidase 9, acid phosphatase 10, alcohol dehydrogenase 11, lipoxygenase 12, PGH synthetase 13, laccase 14, alkaline phosphatase 15, lactate dehydrogenase 16, catalase 17, alcohol dehydrogenase. (From Ref. 38.)...

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