Highly Efficient Synthesis of Novel Fluorine Bearing Quinoline-4-carboxylic Acid and the Related Compounds as Amylolytic Agents

Highly efficient syntheses of novel fluorine bearing quinoline-4-carboxylic acids and the related compounds had been achieved from cyclocondensation of 2-amino-5-fluorophenyl glyoxylic acid 1 with benzoyle asetanilides 2 in boiling DMF, to give the target 3. Decarboxylation of 3 produced 6-fluoro-2-phenyl-3-(substituted amino)-keto-quinolines 4, while that reaction underwent refluxing, afforded 7-fluoro-1-(aryl)-3-phenyl-pyrrolo[3,4-c] quinoline-2,9-diones 5. Structure of the products has been established from their elemental and spectral analysis. All targets exhibited a high to moderate activity against some Aspergillus fungi as amylolytic agents.

Biodiversity of Indigenous Amylolytic and Cellulolytic Bacteria in Sago Waste Product at Susupu, North Moluccas

People at Susupu, North Moluccas prepare the sago （Metroxylon sago） in traditionally way for consumption. The residue of processed sago usually thrown away on the ground, so it was caused pollution. Some amylolytic bacteria species and cellulolytic bacteria species could be founded in sago waste product. The purpose of this research are：1 ） to identify the indigenous amylolytic bacteria species in sago waste product; 2） to identify the indigenous cellulolytic bacteria species in sago waste product; 3） to test the amylum hydrolysis ability of each amylolytic bacteria species; 4） to test the cellulose hydrolysis ability of each cellulolytic bacteria species. This research was conduct at the Microbiology laboratory, Biology Department-FMIPA-State University of Malang and the Microbiology laboratory-Faculty of Medicine-Brawijaya University. The research samples are sago waste product from Susupu, North Moluccas. The samples were grinded and diluted in nutrient broth, and then the suspension was diluted gradually until 10^-10. The suspension was inoculated 0.1 mL each on nutrient agar medium in 37℃ during 1 × 24 h. Each bacteria colony that grows on the medium were isolated and determined to know which one were the amyloliytic bacteria and the cellulolytic bacteria. The amylum hydrolysis index of each amylolytic bacteria species were counted as well as the cellulose hydrolysis index of each cellulolytic bacteria species. Each amylolytic bacteria and cellulolytic bacteria species were identified. This research result shows that： 1） there are 5 indigenous amylolytic bacteria species, i.e., Bacillus mycoides, Bacillus cereus, Bacillus licheniformis, Bacillus alvei and Serratia liquefaciens; 2） there are 4 indigenous cellulolytic bacteria species, i.e., Serratia liquefaciens, Acinetobacter iwofii, Bacillus licheniformis and Bacillus cereus; 3） Serratia liquefaciens has the highest amylum hydrolysis index, i.e., 3.08; 4） Acinetobacter iwoffii has the highest cellulose hydrolysis index, i.e., 2.01.

Isolation of Filamentous Fungi from the Caatinga Region and Production of Amylolytic Enzymes of Great Industrial Interest

The industrial importance and the high cost of the commercial amylase require the study of microorganisms that produce these enzymes. For this reason, the objective of this work was to isolate filamentous fungi from a region of the caatinga and evaluate their potential for the production of amylase. Four soil samples were collected from a deactivated dump located in the city of Diamantina, MG, in a region of the caatinga. The analysis of amylolytic production in a submerged medium at the ideal temperature of each microorganism was performed using the saccharification method, and the reducing sugars formed were quantified by DNS. Fourteen filamentous fungi were isolated, which had different morphological aspects. Regarding amylase production, a mean activity of 0.477 U·mL-1 was obtained with the isolates I 1.2.1 and I 4.4.1. These results bring important information regarding the biodiversity of the caatinga, in addition to the isolation of microorganisms that can be used as biological machinery to obtain metabolites with high biotechnological and industrial potential.

Biodiversity and Enzyme Activity of Indigenous Cellulolytic and Amylolytic Bacteria in Decayed Mangrove Stem Waste Product at Waai Seashore, Ambon Island

Mangrove （Sonneratia spp.） could be found at Waai seashore, Ambon island. The remainder of the mangrove stem will be decayed and become the waste product. Some indigenous bacteria species that live in the decayed mangrove stem waste product have cellulolytic and amylolytic characters. This research was done to： （1） identify and determine the cellulolytic bacteria species; （2） identify and determine the amylolytic bacteria species; （3） determine the cellulolytic bacteria species that have the highest cellulase activity; （4） determine the amylolytic bacteria species that have the highest amylase activity. This research was conducted at the Microbiology Laboratory, Department of Biology, State University of Malang, Microbiology Laboratory, Faculty of Medicine, Brawijaya University and Chemistry Laboratory, Muhammadiyah Malang University. In the experiment, 25 g sample were grinded and diluted in 225 mL nutrient broth to get 101 suspension. Then the suspension was diluted gradually until 101~. The suspension was inoculated on nutrient agar medium with 0.1 mL each, and incubated in 37 ~C during 24 h. Each bacteria colony was isolated and identified to know whether it was cellulolytic or amylolytic bacteria. Afterward, the cellulase activity as well as the amylase activity was analyzed. The research results show that： （1） there are four cellulolytic bacteria species, i.e., Micrococcus luteus, Bacillus pumilus, Planococcus citreus and Bacillus cereus; （2） there are four amylolytic bacteria species, i.e., Bacillus firmus, Nitrobacter sp., Bacillus mycoides and Pseudomonas aeruginosa; （3） Bacillus cereus has the highest cellulase activity; （4） Nitrobacter sp. has the highest amylase activity.

novel amylolytic enzyme from Palaeococcus ferrophilus with malto-oligosaccharide forming ability belonging to subfamily GH13_20

By heterologous expression of a gene from Palaeococcus ferrophilus,a novel recombinant enzyme designated AMPf was obtained and proved to be an amylolytic enzyme with unique catalytic characteristics.The optimal temperature and pH of AMPf were 50℃ and 7.0 respectively.Although sequence analysis and acarbose hydrolysis ability indicated that AMPf belongs to the subfamily GH13_20,interestingly,this enzyme hardly acts on cyclodextrins and pullulan distinguishing it from most enzymes in this subfamily.AMPf hydrolyzes starches to glucose,maltose,maltotriose,and maltotetrose as main products.AMPf mainly liberates glucose from starch with the concentration of 1%(w/v),while it shows malto-oligosaccharide forming ability with higher starch concentration of 4%(w/v).Also,the 4,6-ethylidene-4-nitrophenyl-maltoheptaose hydrolysis ability further indicates the unique combination endo-acting and glucose releasing exo-acting activtity of AMPf.AMPf could utilize maltose and maltotriose to produce malto-oligosaccharides by transglycosylation activity.It was proven AMPf has application protential in malto-oligosaccharides production.

Inter-Conversion of Carbohydrate Reserves from Pollen Maturation to Rehydration in a Chili Pepper

Carbohydrate metabolism is critical for male fertility. Carbohydrate reserves (soluble sugars and starch) were quantified in three advanced stages of pollen development, from the final maturation to rehydration, in a chili pepper cultivar (Capsicum annuum L. cv. Calypso Red) with partially dehydrated pollen, to define possible carbohydrate inter-conversions. At the same time, the activity of soluble enzymes involved in sucrolysis and amylolysis were quantified to reveal potential metabolic pathways. The carbohydrates found (sucrose, glucose, fructose, and starch) apparently inter-convert. There would be a close relation between enzymatic activities and substrates, and the carbohydrate reserves could be key factors on the regulation of enzymatic activities. All the enzymes tested were active, although the relevance of each one changed along pollen maturation and rehydration, defining different profiles of enzymatic activities for each stage. Some usually neglected enzymes (e.g. soluble neutral invertase) have shown an important role in the stages analyzed, suggesting alternative processes to evaluate in the studies of male fertility control.

Sweet Potato Starch as a Carbon Source for Growth and Glucoamylase Production from <i>Aspergillus niger</i>

A mesophilic strain of Aspergillus niger isolated from cassava effluent samples produced extracellular glucoamylase in submerged culture containing 2% (w/v) soluble or sweet potato starch. On soluble starch medium the maximum glucoamylase activity in the culture filtrate was 9.40 U/mg compared to 8.24 U/mg on sweet potato starch culture filtrate. The mycelial dry weight for both media was 494 and 418 mg respectively. The maximum glucoamylase activity was obtained at a growth temperature of 40&deg;C and pH 4.5. The implication is that the bioprocess of utilizing sweet potato starch in the culture is attractive due to its relatively cheaper availability in Nigeria, making it even more favorable when economics is considered.