Production and application of α-amylase from indigenous fungalstrain Aspergillus luchuensis bs1

@#Aim:The study aimed at isolation, screening, optimization and partial purification of α-amylase and evaluating its desizing efficiency in textile industry. Methodology and results:The AF01 showed the highest α-amylase activity of 128 KU. This isolate was identified asAspergillus luchuensisstrain bs1 using 18S rRNA gene sequencing. The process parameters were screened by employing Plackett-Burman Design (PBD) with seven variables and followed by Box-Behnken Design with three positively influencing factors. The investigation revealed that the maximum α-amylase production (192KU) at medium pH 5.6, starch 3% (w/v) and sodium nitrate 0.5% (w/v). The partial purification of α-amylase was done by acetone precipitation and it resulted in 6.1 fold purification. Partially purified α-amylase recorded optimum activity at pH 5.5, 60 min of contact time, temperature stability at 60°C and 93%specificity to potato starch. The desized cotton fabric showed 9.5% weight loss, 5 sec of absorbency time and 8 rating in Tegewa analysis.Conclusion, significance and impact of study: The study proposes a novel indigenous fungal strain having ability to produce alpha amylase and an enzyme preparation for desizing sized cotton fabric in minimal concentration.

Genome Shuffling of Mangrove Endophytic Aspergillus luchuensis MERV10 for Improving the Cholesterol-Lowering Agent Lovastatin under Solid State Fermentation

In the screening of marine mangrove derived fungi for lovastatin productivity, endophytic Aspergillus luchuensis MERV10 exhibited the highest lovastatin productivity (9.5 mg/gds) in solid state fermentation (SSF) using rice bran. Aspergillus luchuensis MERV10 was used as the parental strain in which to induce genetic variabilities after application of different mixtures as well as doses of mutagens followed by three successive rounds of genome shuffling. Four potent mutants, UN6, UN28, NE11, and NE23, with lovastatin productivity equal to 2.0-, 2.11-, 1.95-, and 2.11-fold higher than the parental strain, respectively, were applied for three rounds of genome shuffling as the initial mutants. Four hereditarily stable recombinants (F3/3, F3/7, F3/9, and F3/13) were obtained with lovastatin productivity equal to 50.8, 57.0, 49.7, and 51.0 mg/gds, respectively. Recombinant strain F3/7 yielded 57.0 mg/gds of lovastatin, which is 6-fold and 2.85-fold higher, respectively, than the initial parental strain and the highest mutants UN28 and NE23. It was therefore selected for the optimization of lovastatin production through improvement of SSF parameters. Lovastatin productivity was increased 32-fold through strain improvement methods, including mutations and three successive rounds of genome shuffling followed by optimizing SSF factors.

Analysis of the MAT1-1 and MAT1-2 Gene Ratio in Black Koji Molds Isolated from Meju

Aspergillus luchuensis is known as an industrially important fungal species used for making fermented foods such as awamori and shochu in Japan, makgeolli and Meju in Korea, and Pu-erh tea in China. Nonetheless, this species has not yet been widely studied regarding mating-type genes. In this study, we examined the MAT1-1 and MAT1-2 gene ratio in black koji molds (A. luchuensis, Aspergillus niger, and Aspergillus tubingensis) and in Aspergillus welwitschiae isolated from Meju, a fermented soybean starting material for traditional soy sauce and soybean paste in Korea. The number of strains with the MAT1-1 locus was 2 of 23 (A. luchuensis), 6 of 13 (A. tubingensis), 21 of 28 (A. niger), and 5 of 10 (A. welwitschiae). Fungal species A. tubingensis and A. welwitschiae showed a 1 : 1 ratio of MAT1-1 and MAT1-2 mating-type loci. In contrast, A. luchuensis revealed predominance of MAT1-2 (91.3%) and A. niger of MAT1-1 (75%). We isolated and identified 2 A. luchuensis MAT1-1 strains from Meju, although all strains for making shochu in Japan are of the MAT1-2 type. These strains may be a good resource for breeding of A. luchuensis to be used in the Asian fermented-food industry.