Purification and characterization of a milk clotting protease from Rhizomucor miehei

  title={Purification and characterization of a milk clotting protease from Rhizomucor miehei},
  author={S. Preetha and Rathanam Boopathy},
  journal={World Journal of Microbiology and Biotechnology},
Benzamidine, an inhibitor of serine proteases, was used as an affinity ligand for the purification of aspartyl protease from culture filtrate of Rhizomucor miehei. The two step purification protocol (ion-exchange and affinity chromatography) resulted in a homogenous enzyme preparation with seven-fold purification and a final recovery of 22%. The purified enzyme was free of brown pigmentation, a factor inherently associated with the enzyme; it was stable and active at acidic pH (optimum pH 4.1… Expand
Purification and Characterization of Milk-clotting Enzyme from Solanum dubium Seeds
This research was carried out to purify and characterize a milk-clotting enzyme from Solanum dubium seeds and to investigate the efficiency of the purified enzyme as a rennet substitute in cheeseExpand
Purification and properties of a milk-clotting enzyme produced by Bacillus amyloliquefaciens D4
The milk-clotting enzyme from Bacillus amyloliquefaciens D4 was purified to 17.2-fold with 20% recovery by precipitation in ammonium sulfate and ion-exchange chromatography. The molecular mass of theExpand
Purification and characterization of a milk-clotting protease from Mucor pusillus: Method comparison
Crude enzymatic extract obtained from five fermentations (300 g of wheat bran) was characterized by a clotting activity of 0.34 ± 0.08 UP/ml with a strength ratio of 1/1: 200. The comparative studyExpand
Production and characterization of a milk-clotting enzyme from Aspergillus oryzae MTCC 5341
High ratio of milk clotting to proteolytic activity coupled with low thermal stability strengthens the potential usefulness of milk-clotting enzyme of A. oryzae MTCC 5341 as a substitute for calf rennet in cheese manufacturing. Expand
Production and characterization of milk-clotting enzyme from Bacillus amyloliquefaciens JNU002 by submerged fermentation
This work described a strain Bacillus amyloliquefaciens JNU002 that screened from wheat bran that has a promising characterization and was stable at pH 4–6 and below 40 °C, and this was convenient for storage and transportation. Expand
Extracellular acid protease from Rhizopus oryzae: purification and characterization
Abstract Extracellular aspartate protease from Rhizopus oryzae was purified 91 times with 26% recovery using (NH 4 ) 2 SO 4 fractionation, ion-exchange and size-exclusion chromatographic techniques.Expand
Production and Purification of Fungal Milk Clotting Enzyme from Aspergillus candidus
The search for rennet substitutes such as microbial rennet has increased fold due to increase in the demand for cheese products. Microbial rennet covers about one-third of the cheese consumptionExpand
Extracellular protease from Mucor pusillus: purification and characterization
Extracellular protease from Mucor pusillus was purified 18-fold with 7.56% recovery by ion-exchange chromatography and gel filtration. The enzyme was found to be monomeric in nature, having aExpand
Religiosin B, a milk-clotting serine protease from Ficus religiosa
Abstract A novel milk-clotting serine protease, named religiosin B, is purified from Ficus religiosa . The molecular mass of the protein is 63,000 with p I value of pH 7.6. The proteolytic activityExpand
A novel aspartic protease from Rhizomucor miehei expressed in Pichia pastoris and its application on meat tenderization and preparation of turtle peptides.
A novel aspartic protease gene (RmproA) was cloned from the thermophilic fungus Rhizomucor miehei CAU432 and expressed in Pichia pastoris, which may be a potential candidate for several industrial applications. Expand


Characterization of two aspartyl proteinases from a commercial fungal (Mucor miehei) rennet
Gel permeation chromatography and isoelectric focusing of the proteins from a commercial fungal rennet resulted in the isolation and purification of two aspartyl proteinases, AP1 and AP2, which were found to have different physicochemical properties. Expand
Dialysis, preparative isoelectric focusing (IEF) and gel filtration were employed to obtain final enrichments of 2.2-fold and 1.4-fold for Mucor miehei (CBS 360.75) proteinase (MMP) and M. pusillusExpand
Crystalline Milk-Clotting Protease from Mucor Miehei and Some of its Properties
The effects of inhibitors indicate that this clotting enzyme is not metal dependent and does not have serine or SH active groups, and the enzyme is most stable at acid pH. Expand
Affinity purification and properties of cathepsin-E-like acid proteinase from rat spleen.
A unique acid proteinase different from cathepsin D was purified from rat spleen by a method involving precipitation at pH 3.5, affinity chromatography on pepstatin-Sepharose 4B and concanavalinExpand
Thermal Inactivation of Residual Milk Clotting Enzymes in Whey
Abstract Cheddar cheese whey from milk clotted with six milk clotting enzymes ( Mucor miehei, Mucor pusillus var Lindt , and Endothia parasitica proteases; calf rennet, porcine pepsin, andExpand
Rapid isolation of microbial milk-clotting enzymes by N-acetyl (or N-isobutyryl)-pepstatin--Aminohexylagarose.
Two affinity Columns were prepared and found to be effective for the rapid purification of microbial milk-clotting enzymes from Mucor miehei and Endothia parasitica and were purified to a homogeneous state from commercial preparations by only one step of each of the affinity chromatographies. Expand
Influence of culture conditions on the production of milk-clotting enzyme from Rhizomucor
Solid-state fermentation on wheat bran for 5 days at room temperature gave optimal production for enzyme by Rhizomucor miehei and R. pusillus to determine the quality of milk-clotting enzyme. Expand
Immunological identification of milk-clotting enzymes
The 6 most widely used milk-clotting enzymes, i.e. chymosin, bovine and porcine pepsins and proteinases from Mucor miehei, M. pusillus and Endothia parasitica , have been purified and used to prepareExpand
Rapid and Large Scale Isolation of Chymosin (Rennin) by Pepstatin-aminohexylagarose
A rapid and large scale isolation of chymosin was achieved by an affinity column including pepstatin-aminohexylagarose with a column of DEAE-cellulose. The affinity column with 3 ml wet gel made itExpand
Acid proteases from species of Mucormii. partial characterization of the acid protease produced by a strain of Mucor miehei isolated in Cuba.
It was suggested that the two enzymes are similar but not identical, and this conclusion was reinforced by an analysis of circular-dichroism spectra. Expand