Validation of scale-up production of microbial rennet

The applicability of coconut paring cake as substrate for solid substrate fermentation. Analysis of the locally produced cheese coagulants (granulated and liquid coagulants) produced from Rhizopus chinensis showed that the coagulants conformed to specifications set by JEPCA [] in the Microbiological Guidelines for specification and Considerations for Enzyme Preparations Used in Food Processing in terms of coliform. E. coli, S. aureus and Salmonella. These specific pathogenic microorganisms were absent in 25g/mL of the samples. It was noted that heterotrophic and aerobic plate counts for both coagulants exceeded the guideline's acceptable values (greater 50,000 CFU/g or mL). Yeast and mold counts for both coagulants also failed to meet acceptable counts specified for commercially available coagulant. Environmental monitoring, particularly the assessment of air quality inside the processing area and the safety cabinet, showed non-conformance to the established standards. Interestingly, the cleaning procedure and practices employed in cleaning contact surfaces in the processing area were effective, as indicated by the attained hygiene status of-PASSI or less than or equal to 100 RLU using the ATP biolumino for the production of microbial rennet was validated through the kinetics study made on the growth Rhizopus chinensis Saito BIOTECH 3273. Growth kinetics of R. chinensis was evaluated using substrate-independent kinetic models namely linear, exponential, logistic and two-phase. Among the four equations used, only the linear model cannot describe the growth of R. chinensis due to a negative value of initial biomass (-0.136%). The logistic model has the highest specific growth rate (0.414/h), closest predicted initial biomass concentration (0.1155%) to experimental data (0.083%), and the least SSE (0.126). It is therefore concluded that the logistic model best described the growth of R. chinensis BIOTECH 3273 using sold substrate fermentation of coconut paring cake. Optimization study was conducted to determine the most suitable conditions for the microbial rennet production of Rhizopus chinensis BIOTECH 3273 in a solid substrate fermentation system of coconut paring cake. In the scale of production of microbial rennet, the target volume for liquid rennet was easily attained. Microbial evaluation of raw materials in the raw materials and the process was done before of the complete installation of the equipment needed in the processing area for rennet production. The possibility of utilizing the fermentation by-product (fermented coconut paring meal or FCPM) as feed ingredient for broilers, a total of 300 day-old chicks were subjected for 38-day feeding trial. Under the conditions which the study was conducted, it can be concluded that the addition of FCPM in the broiler ration had reduced the growth performance of broilers indicating poor feed utilization. DNA sequencing was done to determine the amino acid sequence of the rennet protein. Under the specified PCR conditions, the primer set RCAP II_F2/RCAP II_R2 successfully amplified a single DNA and of the expected product length in both the CBS Type Strain (standard organism) and BIOTECH 3273. With primers set RCAP II_R5, the expected product was faintly amplified only in CBS. Production of rennet casein from skimmed milk was studied and was used in the production of imitation cheese to improve fat and water binding, texture and matrix formation. Numerous trainings were conducted in different parts of the country to demonstrate the application of microbial rennet in cheese making. Furthermore, several technology forums and food expo were attended to promote microbial rennet to dairy processors and potential technology takers.