Abstract A thermophilic Bacillus strain ASU7 was isolated from soil sample collected from Assiut governorate in Upper Egypt on latex rubber-containing medium at 45 °C. Genetically, the 16S bacterial ribosomal RNA gene of the strain ASU7 was amplified by the polymerase chain reaction (PCR) and sequenced. The sequence of the PCR product was compared with known 16S rRNA gene sequences in the GenBank database. Based on phylogenetic analyses, strain ASU7 was identified as Bacillusamyloliquefaciens. The strain was able to utilize Ficus elastica rubber latex as a sole source for carbon and energy. The ability for degradation was determined by measuring the increase in protein content of bacterium (mg/g dry wt), reduction in molecular weight (g/mol), and inherent viscosity (dl/g) of the latex. Moreover, the degradation was also confirmed by observing the growth of bacterium and formation of aldehyde or keto group using scanning electron microscopy (SEM) and shiff’s reagent, respectively.

Abstract A thermophilic Bacillus strain ASU7 was isolated from soil sample collected from Assiut governorate in Upper Egypt on latex rubber-containing medium at 45 °C. Genetically, the 16S bacterial ribosomal RNA gene of the strain ASU7 was amplified by the polymerase chain reaction (PCR) and sequenced. The sequence of the PCR product was compared with known 16S rRNA gene sequences in the GenBank database. Based on phylogenetic analyses, strain ASU7 was identified as Bacillusamyloliquefaciens. The strain was able to utilize Ficus elastica rubber latex as a sole source for carbon and energy. The ability for degradation was determined by measuring the increase in protein content of bacterium (mg/g dry wt), reduction in molecular weight (g/mol), and inherent viscosity (dl/g) of the latex. Moreover, the degradation was also confirmed by observing the growth of bacterium and formation of aldehyde or keto group using scanning electron microscopy (SEM) and shiff’s reagent, respectively.

Abstract A thermophilic Bacillus strain ASU7 was isolated from soil sample collected from Assiut governorate in Upper Egypt on latex rubber-containing medium at 45 °C. Genetically, the 16S bacterial ribosomal RNA gene of the strain ASU7 was amplified by the polymerase chain reaction (PCR) and sequenced. The sequence of the PCR product was compared with known 16S rRNA gene sequences in the GenBank database. Based on phylogenetic analyses, strain ASU7 was identified as Bacillusamyloliquefaciens. The strain was able to utilize Ficus elastica rubber latex as a sole source for carbon and energy. The ability for degradation was determined by measuring the increase in protein content of bacterium (mg/g dry wt), reduction in molecular weight (g/mol), and inherent viscosity (dl/g) of the latex. Moreover, the degradation was also confirmed by observing the growth of bacterium and formation of aldehyde or keto group using scanning electron microscopy (SEM) and shiff’s reagent, respectively.

Abstract Bioaugmentation for the removal of polyaromatic hydrocarbons (PAHs) from wastewater using bacteria and yeasts is considered environment-friendly and a cost-effective technique. The effectiveness of this biodegradation system depends on the stability of inoculated microorganisms and the availability of nutrients. This study is aimed to investigate the removal of high molecular weight (HMW)-PAHs from biologically treated produced water using different biological systems. Three systems, inoculated with activated sludge (AS), the mixture of five yeast strains (MY), and the mixture of AS and the five yeast strains (SY), respectively, were constructed, and their performance for the removal of HMW-PAHs was compared over 10 weeks. The effluent of the biologically treated produced water from an oilfield was used as the influent after chrysene and benzo(a)pyrene were spiked as HMW-PAHs. Polymerase chain reaction-based denaturing gradient gel electrophoresis (PCR-DGGE) and fluorescent in situ hybridization (FISH) techniques were used to examine the changes in the structures and abundances of the bacterial and yeast communities in these three systems. Only SY and MY systems were capable to remove chrysene (90.7 % and 98.5 %, respectively) and benzo(a)pyrene (80.7 % and 95.2 %, respectively). PCR-DGGE analysis confirmed that all of the five yeast strains inoculated remained in the SY and MY systems, while FISH results showed that the relative abundance of yeast in the SY and MY systems (10.6 % to 21.9 %, respectively) were significantly higher than AS system (2.3 % to 7.8 %, respectively). The relative abundances of the catechol 2,3-dioxygenase (C23O) indicated that the copy number ratios of benzene ring cleavage gene C23O in the yeast amended systems were much higher than that in the AS system. In this study, all of the three systems were effective in removing the low molecular weight (LMW)-PAHs, while HMW-PAHs including chrysene and benzo(a)pyrene were efficiently removed by MY and SY systems, not by AS system. The high HMW-PAHs removal in the MY and SY bioaugmentation systems possibly attributed to the inoculation of the mixed yeast culture. By combining the PCR-DGGE results with the FISH analyses, it was found that yeast probably consisting mainly of the five inoculated strains inhabited in the two bioaugmentation systems as a dominant population. The relatively higher performance of the SY system might be attributed to the suspended growth type which permitted a more efficient contact between microbial cells and contaminants. The bioaugmentation systems (SY and MY) were successfully established by inoculating with five nonindigenous yeast strains and demonstrated high performance in removal of HMW-PAHs.

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Abstract : Bio-ethanol has been promoted as alternative fuel because it is renewable fuel and environmental friendly compared with petroleum products. In this study ethanol production by a yeast isolate, designated as YB2 isolated from Kenyan dairy industry, in whey waste was examined. The isolate was initially identified as Pichia cactophila using RFLP of PCR-amplified internal transcribed spacers of rDNA (ITS1-5.8S-ITS2). Where size of the PCR products (450 bp), and the restriction analyses with two restriction enzymes HaeIII and HinfI yielded fragments with 450 bp and 200+250 bp respectively. The variable D1/D2 domain of the 26S rDNA of the isolate was amplified by PCR and sequenced. The sequences were compared with known 26S rDNA sequences in the GenBank database. Results of 26S rRNA gene confirmed that the isolate was highly related to P. cactophila with similarity 100%. Phylogenetic analysis shows that YB2 shared a one cluster with P. cactophila. The fermentative performance of the strain YB2 on cheese whey to produce ethanol was evaluated at different parameters such as incubation temperature, initial pH, whey sugar concentrations, and yeast concentrations. Maximum ethanol produced by strain YB2 was achieved at pH 4.5 and 35°C. To our knowledge, there are no reports in literature on utilization of cheese whey by P. cactophila for ethanol production. This was therefore, the first time the P. cactophila was used to produce ethanol from cheese whey.

Abstract The utilization of agro-industrial wastes such as whey as raw materials for the production of bio-ethanol is gaining importance as a result of the attractiveness of renewable fuel alternatives due to exhaustion of fossil fuel sources coupled with the positive impact to the environment. Here, we report the isolation of two Kluyveromyces spp. designated as BM4 and P41, able to produce ethanol as main fermentation product from fermenting whey. Three different molecular biological approaches including, the RFLP analysis of the 5.8S-ITS rDNA, the sequence of the 5.8S-ITS rDNA region and the sequence of the D1/D2 domain of the 26S rRNA gene were applied for accurate identification. While RFLP analysis of 5.8S-ITS region failed to accurate the differentiation between the two species, sequencing of this region and D1/D2 region of the 26S rRNA gene verified the identification. PCR amplification and sequence analysis of 5.8S-ITS rRNA and D1/D2 domain of the 26S rRNA genes revealed that the isolates BM4 and P41 were highly related to Kluyveromyces marxianus and Kluyveromyces lactis with homology of 99% for both. In addition, phylogenetic analysis indicated that both BM4 and P41 shared a cluster with K. marxianus and K. lactis, respectively. The fermentative performance of both strains on cheese whey to produce ethanol was evaluated at different parameters such as incubation temperature, initial pH, whey sugar concentrations, and yeast concentrations. Results show that the maximum ethanol productions achieved at pH 4.5 and 35 °C were 5.52% and 5.05% for K. marxianus and K. lactis, respectively. Our results demonstrated that K. marxianus and K. Lactis could be recommended for cheese whey bioremediation in the environment and produce renewable biofuel.

Abstract: Cheese whey is a dairy industry waste have high lactose concentration, which case environmental problem. Therefore biofuel production from cheese whey become an interest as renewable energy. In this study 28 yeasts were isolated from Kenyan cheese whey and characterized for lactose fermentation. Out of them one isolate, designated as BM9 had high efficiency for lactose fermentation was selected. For genetically identification PCR amplification of both internal transcribed spacer (ITS) and 5.8 rDNA region (ITS1-5.8S-ITS2) and the variable D1/D2 domain of the 26S rRNA gene were used. RFLP of the PCR-amplified ITS1-5.8S-ITS2 region (750 bp) using two restriction enzymes HaeIII and HinfI yielded fragments with 650 + 80 bp and 250 + 190 + 120 + 80 bp respectively witch identified the isolate as Kluyveromyces marxianus. 26S rRNA of the isolate was sequenced and compared with known 26S rRNA sequences in the GenBank database. Results of 26S rRNA gene confirmed that the isolate was highly related to K. marxianus with similarity 99%. Phylogenetic analysis show that BM9 shared a one cluster with K. marxianus. The fermentative performance of the strain BM9 on cheese whey to produce bio-ethanol at different parameters such as incubation temperature, initial pH, whey sugar concentrations, and yeast concentrations was evaluated.