Use of a new milk-clotting protease from Thermomucor indicae-seudaticae N31 as coagulant and changes during ripening of Prato cheese

Prato cheeses were manufactured using coagulant from Thermomucor indicae-seudaticae N31 or a commercial coagulant. Cheeses were characterised using the following analysis: yield; fat; acidity; moisture; ash; salt; pH; total nitrogen; total protein; NS-pH 4.6/NT100; NS-TCA 12%/NT100; casein electrophoresis; and RP-HPLC. The results were statistically analysed and revealed that the proteolytic indices were not significantly different throughout the 60 days of ripening of cheeses made with either coagulant. Even though there were some quantitative differences in the peptide profile of cheeses, the enzyme from T. indicae-seudaticae N31 was used in the production of good quality Prato cheese without having to change the established technological parameters of the process. © 2011 Elsevier Ltd. All rights reserved.

Production and Characterization of a Milk-clotting Protease Produced in Submerged Fermentation by the Thermophilic Fungus Thermomucor indicae-seudaticae N31

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Production and characterization of a milk-clotting protease in the crude enzymatic extract from the newly isolated Thermomucor indicae-seudaticae N31 (Milk-clotting protease from the newly isolated Thermomucor indicae-seudaticae N31)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Purification, characterization and crystallization of Jararacussin-I, a fibrinogen-clotting enzyme isolated from the venom of Bothrops jararacussu

A fibrinogen-clotting enzyme, Jararacussin-I, was purified from the venom of Bothrops jararacussu by a combination of ion exchange chromatography using Resource 15S resin and affinity chromatography using Benzamidine Sepharose 6B resin. Jararacussin-I displays a molecular mass of 28 kDa as estimated by sodium dodecyl sulphate-PAGE and possesses an isoetectric point of 5.0. The coagulant specific activity of the enzyme was determined to be 45.8 NIH U/mg using bovine fibrinogen as the substrate and the esterase specific activity was determined to be 258.7 U/mg. The protease inhibitors, benzamidine and DTT inhibited the esterase specific activity by 72.4 and 69.7%, respectively. The optimal temperature and pH for the degradation of both chains of fibrinogen and esterase specific activity were determined to be 37 degreesC and 7.4-8.0, respectively. The enzyme was inactivated at both 4 and 75 T. Single crystals of Jararacussin-I were obtained and complete three-dimensional X-ray diffraction data was collected at the Brazilian National Synchrotron Source (LNLS) to a resolution of 2.4 Angstrom. (C) 2002 Published by Elsevier B.V. Ltd.

Produção e caracterização da protease coagulante obtida por fermentação submersa a partir do fungo termofílico Thermomucor indicae-seudaticae N31

Pós-graduação em Engenharia e Ciência de Alimentos - IBILCE

Produção, purificação e caracterização da protease de Thermomucor indicae seudaticae N31 e avaliação de sua aplicação na fabricação do queijo maturado

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Caracterização bioquímica e funcional da ação da peptidase de Thermomucor indicae-seudaticae N31 sobre a caseína

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Yield, changes in proteolysis, and sensory quality of Prato cheese produced with different coagulants

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Biochemical and functional properties of a thrombin-like enzyme isolated from Bothrops pauloensis snake venom

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Milk fatty acid characterization and genetic parameter estimates for milk conjugated linoleic acid in buffaloes

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Detection of nontuberculous mycobacteria from water buffalo raw milk in Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Time-dependent alterations of soluble and cellular components in human milk

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Activity of Delta(9)-desaturase enzyme in mammary gland of lactating buffaloes

The objective of this research was to measure the activity of e-desaturase enzyme in lactating buffaloes. Data from forty lactating Murrah-crossbred buffaloes were collected on five commercial farms located at Sarapui and Pilar do Sul, São Paulo-Brazil. A field survey was done from April to November 2002. In four farms, buffaloes were fed with wet brewers grains (primary concentrate). Only one farm (Farm 4) offered pasture and corn silage. Monthly milk samples were collected and stored at -20 degrees C until analyzed for fatty acid composition. The Delta(9)-desaturase activity was measured using an indirect method (myristoleic and myristic acids ration - C(14:1c9)/C(14:0)). The higher C(14:1c9)/C(14:0) rate was verified on Farm 4 (0.092). The C(14:1c9)/C(14:0) ratio were 0.064 to Farm 1; 0.065 to Farm 2; 0.062 to Farm 3 and 0.065 to Farm 5. The C(17:1)/C(17:0), C(18:1c9)/C(18:0) and C(18:2c9t11)/C(18:1t11) ratios were also affected. The Farm 4 showed higher value for all ratios. Therefore, in lactating buffaloes grazing pasture the Delta(9)-desaturase activity could be enhanced.

An enzyme-linked immunosorbent assay (ELISA) for detection of antibodies against Toxocara vitulorum in water buffaloes

Toxocara vitulorum, a parasite of the small intestine of cattle and water buffaloes, is mainly acquired by calves via the colostrum/milk from infected cows. To understand the development of immune responses in calves, antibody levels to a soluble extract antigen (Ex) from T. vitulorum infective larvae were measured by an indirect ELISA with sera of 15 buffalo calves, which were sampled every 15 days for the first 180 days after birth and 9 buffalo cows during the perinatal period. From all serum samples examined during the first 180 days, antibody level was lowest and highest in calves at 1 day of age before and after suckling colostrum, respectively, suggesting that the origin of antibodies was the colostrum. Immediately after birth, antibody levels in suckled calves remained at high levels until day 15, began to decrease to lower levels between 15 and 30 days and remained relatively stable until 120 days. By comparing the immune responses of these animals with their parasitological status it was considered possible to determine if passively acquired or actively produced antibodies provided protection against the infection. High numbers of T. vitulorum eggs in the feces between 30 and 60 days indicated that passively acquired antibodies did not provide protection against the infection, at least during these first days, and the maximum fecal egg counts during 30-45 days were coincident with decreased antibody levels. Between 60 and 120 days, when serum antibodies were detected at reduced, but stable levels, adult nematodes were expelled from the intestines and no more T. vitulorum eggs were found, suggesting development of acquired resistance. However, the potential and functional protective role of the antibodies against T. vitulorum infection and the process of self-cure requires further investigation. (C) 2001 Elsevier B.V. B.V. All rights reserved.

Use of PCR-RFLP (Polymerase Chain Reaction-Restricted Fragment Length Polymorphism) in the gene of the enzyme Stearoyl-CoA-Desaturase in Bubalus bubalis

The milk is an important food because it contents Conjugated Linoleic Acids (CIA). These fatty acids are synthesized in mammary gland under action of the enzyme Stearoyl CoA-Desaturase (SCD) and have showed some positive effects in human disease prevention and treatments. A variation of CLA in milk fat exists and can be partially explained by the different levels of expression of SCD. The aim was to study part of the encoding regions of SCD's gene using PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism). Genomic DNA was extracted from lactating Murrah females. After this, PCR reactions were made by using primers Z (sic) (sic) D1 that encloses exon I, II and intron I. The fragments amplified are composed by 938 pb. Then, RFLP techniques were applied in the fragments using the restriction enzymes Pst I and Sma I. The enzyme Pst I has generated fragments of 788pb and 150bp and the Sma I has generated fragments of 693pb and 245pb. All the animals showed the same migration standard for both enzymes, characterizing a genetic monomorphism for this region of SCD gene. The analysis determined that there aren't genetic differences between these animals in the studied regions by using Pst I and Sma I enzymes.