Influence of germinated brown rice-based flour modified by MAse on type 2 diabetic mice and HepG2 cell cytotoxic capacity.

This study aimed to discover whether using maltogenic amylase (MAse) to modify starch in germinated brown rice flour may enhance slow digestion starch and release more bioactive compounds (BCs) content. To achieve this aim, the starch was modified with four levels of MAse (0 U, 133 U, 266 U and 399 U MAse/g flour) for 1 hr at pH 5 and then spray-dried to make modified flour. The biochemical impacts of the products were then accessed in normal and type 2 diabetic mice for 4 weeks. The result showed that when the starch was modified by MAse 266 U/g, a significant reduction of rapidly digested starch to 22.35% from 61.56%, an increase in slowly digested starch to 33.09% while resistant starch as 2.92% corresponding to the increase of γ-amino butyric acid to 528.1 ± 44.1 mg/L and 120.6 ± 10.9 mg/L of ferulic acid. The extract from modified flour showed very strong cytotoxic activity against HepG2 cell (>80% inhibition). The result in vivo showed that the type-2 diabetic mice fed with this modified product could better improve the stability of the glycemic index. Also, atherosclerotic plaque assessment further supports these findings. The results indicated that BCs released considerably couple with the changes in starch properties caused by MAse enhanced the effectiveness of this product to diabetes as well as positive effect on cytotoxic activity against HepG2 cell.

Taka-amylase A in the conidia of Aspergillus oryzae RIB40.

A study of Taka-amylase A of conidia from Aspergillus oryzae RIB40 was done. During the research, proteins from conidia and germinated conidia were analyzed using SDS-PAGE, 2-D gel electrophoresis, Western blot analysis, MALDI-TOF Mass spectrometry, and native-PAGE combined with activity staining of TAA. The results showed that TAA exists not only in germinated conidia but also in conidia. Some bands representing degraded products of TAA were detected. Conidia, which formed on starch (SCYA), glucose (DCYA), and glycerol (GCYA) plates, contained mature TAA. Only one active band of TAA was detected after native-PAGE activity staining. In addition, TAA activity was detected in cell extracts of conidia using 0.5 M acetate buffer, pH 5.2, as extraction buffer, but was not detected in whole conidia or cell debris. The results indicate that TAA exists in conidia in active form even when starch, glucose, or glycerol is used as carbon source. TAA might belong to a set of basal proteins inside conidia, which helps in imbibition and germination of conidia.

Analysis of secreted proteins during conidial germination of Aspergillus oryzae RIB40.

To broaden our understanding of extracellular proteins of Aspergillus oryzae at the conidial germination stage, analyses of the secreted proteins during germination were carried out. Taka-amylase A (TAA), glucoamylase (GLAA), and aspergillopepsin A (PEPA) were identified as the main products by peptide mass fingerprinting. TAA and PEPA were detected simultaneously with the formation of germ tubes. With the development of germination, the pH of the medium fell from 5.5 to 3.5. The secreted PEPA had a pro-sequence and likely shifted from 42 kDa to 41 kDa below pH 4.6, indicating that the precursor of PEPA was secreted and underwent pH-dependent processing. Furthermore, the 41 kDa protein was trapped by the addition of pepstatin A, the specific inhibitor of PEPA, suggesting that the maturation of pro-PEPA was a stepwise autoprocessing upon acidification of the medium and itself was an intermediate of the processing. It was implied that PEPA plays an important role at the early germination stage.