Statistical enhancement of lipase extracellular production by Bacillus stratosphericus PSP8 in a batch submerged fermentation process.

AIM: The aim of this study was to isolate and identify lipolytic bacteria. Perform a statistical stepwise physicochemical optimization for maximum production of extracellular lipase and its validation in a bioreactor. METHODS AND RESULTS: Several lipolytic bacteria were isolated from petroleum hydrocarbon-polluted soil. The strain expressing the highest lipase activity (47 U ml-1 ) was genetically identified as Gram-positive Bacillus stratosphericus PSP8 (NCBI GenBank accession no. MH120423). The response surface methodology (RSM)-central composite face centre (CCF) design of experiments was performed based on the preselected levels of the studied parameters obtained from the performed one-factor-at-a-time sequential experiments. A second-order polynomial model was predicted and improved the lipase production by approximately 1·6-fold. Preliminary scaling up of the validated optimized process was carried out in a batch 10-l stirred tank bioreactor, applying the optimum predicted operating conditions; pH 6·98, 34·8°C, 2·2 × 106 cells per ml, 200 rev min-1 , 4·82 g l-1 tributyrine concentration, 1% sucrose and 0·1% yeast extract. This yielded 89 U ml-1 at the late log phase of bacterial growth (48 h). Logistic kinetic model effectively characterized the submerged fermentation process, and the maximum specific growth and lipase production rates were estimated to be 0·338 and 0·164 h-1 respectively. CONCLUSIONS: The mesophilic and neutrophilic B. stratosphericus PSP8 isolated from petroleum hydrocarbon-contaminated soil is a proper source of lipase. The closeness of the predicted response with that of the experimental value and the enhancement of lipase productivity in fermenter scale by approximately 1·9-fold, showed that statistically optimized design can be used in order to improve the lipase production to meet the increasing demand. SIGNIFICANCE AND IMPACT OF THE STUDY: The RSM-CCF statistical optimization is useful for optimizing a large number of variables and studying their interactive effects on extracellular lipase production.

ROBERTS SYNDROME: CLINICAL AND CYTOGENETIC STUDIES IN 8 EGYPTIAN PATIENTS AND MOLECULAR STUDIES IN 4 PATIENTS WITH GENOTYPE/PHENOTYPE CORRELATION.

The Roberts syndrome (RBS) is a rare autosomal recessive disorder caused by mutation in ESCO2 gene. Among over 150 reported international cases, 16 cases are Egyptian including the presently reported patients. The current study reports 8 new Egyptian patients from 7 unrelated consanguineous families investigating clinical phenotype as well as cytogenetic changes in all cases and mutational spectrum in 4 cases. Clinical, orodental, cytogenetic and molecular studies were done to investigate genotype/phenotype correlation. Evaluation of the studied 8 patients showed that they all exhibited the main limb and craniofacial features of Roberts syndrome. Cytogenetic studies including centromeric separation and puffing by Giemsa and DAPI stains and for the first time in Egypt analysis for premature centromeric division by FISH showed consistent centromeric separation in all studied cases. Molecular studies of 4 available patients showed that they all have ESCO2 gene mutation. We conclude that RBS has a well-defined clinical spectrum. The cytogenetic changes are due to sister chromatid cohesion defects which lead to mitotic dysfunction. We confirmed previous results of lack of genotype/phenotype correlation. We also confirmed that the severity of limb malformation correlates with craniofacial manifestations. We recommend detailed evaluation of orodental changes for further definition of the phenotype and for proper patient management. We emphasize the need for further studies for the frequency of premature centromeric separation by FISH as a possible indicator of phenotypic severity.

Selenium metabolism in a strain of Fusarium.

Fusarium sp. was isolated from Sinai soil at Egypt. It showed tendency to tolerate high concentrations of selenium in the form of sodium selenite up to 3.5% (w/v). The microscopic examination revealed some morphological distortions. However, the fungus was capable to circumvent the toxic effect of selenium. The fungus possess strong reducing ability as high quantities of elemental selenium were precipitated within the fungal cells as well as on the surface of the fungal hyphae and spores. The presence of selenium increased the cellular contents of carbohydrates, proteins, and lipids. Labeling studies indicate the incorporation of selenite into certain amino acids: selenocysteine and selenocysteic acid. Moreover, the presence of selenium induced the biosynthesis of several types of low molecular weight proteins. The results demonstrated different modes of detoxification of selenium toxicity.