The present study reports the optimization of process parameters for L-glutaminase production  by  Pseudomonas species ALG3. The L-glutaminase –producing bacterium had already been isolated from soil at a location within Chukwumemeka Odumegwu Ojukwu University, Uli in Anambra state. It was identified using cultural, biochemical and molecular characteristics. Optimization of L-glutaminase production was done by assessing the effect of various parameters using mineral salt medium contained in 250 ml flasks. The parameters included various bivalent metals, pH, surfactants, growth promoters, amino acids and fermentation time. The Results showed that the addition of FeSO4 stimulated optimum L-glutaminase yield of 38.36 U/ml by Pseudomonas species ALG3, while the least production was observed with MnSO4. Enhanced L-glutaminase yield of 30.73 U/ml was recorded at pH 8.0, while the lowest enzyme accumulation was observed at pH 9.0. The highest L-glutaminase accumulation of 82.25 U/ml was achieved with Tween 80, while the lowest yield was recorded with stearic acid. Urea, stimulated maximum L- Glutaminase yield of 43.82 U/ml, while minimum accumulation was observed with peptone. Proline stimulated highest L- glutaminase yield of 42.63U/ml, while the lowest yield was observed with tyrosine. Highest L- glutaminase yield (91.88 U/ml) was observed after 72h, the yield decreased thereafter. The results obtained in the study illustrated that the optimization of process parameters, increased appreciably the L-glutaminase yield of Pseudomonas species ALG3. This suggest that Pseudomonas species can be used for large scale production of L- glutaminase, which can offer great potential for applications in Medicine and food industry.

Proteases are one of the most industrially important enzymes, which account for about 60% of total enzyme market. Protease production by submerged fermentation in shake flasks using Bacillus sp. isolated from the soil was studied. Soil samples were collected from different locations within Chukwuemeka Odumegwu Ojukwu University, Uli, Anambra state. The soil samples were serially diluted and inoculated on sterilized skim milk agar plates. The plates were incubated at 30oC for 72 h. A clear zone around the colonies gave an indication of protease-producing bacteria isolates.   The selected protease producers were subsequently used for shake flask fermentation in 50 ml sterile medium. Optimization study was conducted to determine the effect of carbon sources, nitrogen sources, trace elements, agitation rates and pH.  Twenty one bacteria isolates were found to be active protease producers and isolates RS-5 and OS-9 had the highest zone of clearance of 13.5 and 12.1 mm respectively. The result of submerged production of protease by the  bacteria isolates  revealed that the isolates RS-5 and OS-9 accumulated maximum protease yield of 3.23 and 2.71 U/ml respectively. The isolates were Gram positive and endospore formers, and were identified as Bacillus sp. RS-5 and OS-9.The addition of Starch and maltose stimulated optimum protease production of 3.47 and 2.77 U/ml by Bacillus sp. RS-5 and OS-9 respectively. Beef extract enhanced maximum enzyme yield of 3.35 and 2.90 U/ml for Bacillus sp. RS-5 and OS-9 respectively. Maximum protease yield of 3.28 U/ml for Bacillus sp. RS-5 and 2.85 U/ml for Bacillus sp. OS-9 was obtained by the supplementation of 0.4 g/l of FeS04 respectively. The maximum protease yield was observed at agitation rate of 200 rpm for Bacillus sp. RS-5 and 170 rpm for Bacillus sp. OS-9. At pH8, protease accumulation was highest for Bacillus sp. RS-5 and OS-9. The study revealed that the soil harbours some protease-producing bacteria strains and protease production can be greatly enhanced through optimization of process parameters.