The Use of a Proteolytic Enzyme of Streptomyces Griseus (Protease G) in Blood Banking:

Abstract

Proteaae C is a comparatively new proteolytic enzyme obtained as a by-product in the manufacture of streptomycin. In this first study we have found it to be of great value in detecting incomplea anti-Rh and other antibodies present in the rera of prenatal cases. Five per cent saline mapena i m of selected cella were exposed to a 0.1 per cent mlution of protease C, and results were compared with thooe obtained by d n g trypsinizcd and Rdnizcd cells. In general, the results compare favorably with those ubtained with ficin, and this new enzyme is superior to trypoin in titration rtudies. It is predicted that protease G will be a lwful addition to the proteolytic enzymes that are currently used in many blood banking procedures. SINCE 1957 when Morton and Pickles7 first advocated the use of trypsin in the detection of incomplete anti-Rh antibodies, the value of trypsin itself,s. 9.17 and subsequently of papain,3. 5 ficin,l. 2 and bromelin has become well established in Rh antibody screening tests. Although Pirofsky and Mangumla* 13 suggesi that bronielin, in particular, should be employed as a standard crossmatch procedure, it is generally accepted that no one enzyme technic is adequate in itself,6 and most workers employ the indirect Coombs test in all major crossmatches, with the addition of a separate bovine albumin or enzyme method at 37 C.

Hydrolysis of surimi wastewater for production of transglutaminase by Enterobacter sp. C2361 and Providencia sp. C1112

“…Before the hydrolysis, the proteases were determined for apparent activity using casein assay (Anson, 1938;Lowry, Rosebrough, Farr, & Randall, 1951). The hydrolysis reaction was performed by using 0.1 U of enzyme to 1 g of protein substrate (based on Kjeldahl's method) in 200 mM phosphate buffer at the optimal pH and incubated in the shaking water bath at the optimal temperature for each enzyme (Buchanan & Dierich, 1965;Hale, Greer, Trinh, & James, 2005;Jo et al, 2008;Shahidi, Han, & Synowiecki, 1995). The sample was collected at a designated time and the reaction was terminated by heating the mixture at 95°C for 5 min and kept at 4°C.…”

Significance of Rh-sensitization during Pregnancy: Its Relation to a Preventive Programme

“…(1) O, RIR2 cells pretreated with 1% ficin; (2) 0, R5R2 cells pretreated with 0.1% protease G; (3) 0, D--/Dcells pretreated with 1 % ficin ; (4) 0, D--/D--cells pretreated with 0.1 % protease G (Buchanan, 1956;Buchanan and Dierich, 1965); and (5) 0, D--/D-untreated cells (indirect antiglobulin test using an anti-IgG serum). In every case the ficinor protease-G-treated test cells gave the best evidence of early sensitization, and probably all but one case would have been missed if reliance had been placed solely on a saline or albumin indirect antiglobulin test as the main screening procedure.…”

A Novel Human Alloantibody in the Gerbich System

“…The treatment of red cells with neuraminidase, bromelain, ficin, papain, Pronase and trypsin in two-stage techniques has been previously described [10,11]. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of solubilized erythrocyte membranes was performed as de scribed by Anstee et al [4],…”