Optimization of pineapple pulp residue hydrolysis for lipid production by Rhodotorula glutinis TISTR5159 using as biodiesel feedstock.

The higher lipid productivity of Rhodotorula glutinis TISTR5159 was achieved by optimizing the pineapple pulp hydrolysis for releasing the high sugars content. The sequential simplex method operated by varied; solid-to-liquid ratio, sulfuric acid concentration, temperature, and hydrolysis time were successfully applied and the highest sugar content (83.2 g/L) evaluated at a solid-to-liquid ratio of 1:10.8, 3.2% sulfuric acid, 105 °C for 13.9 min. Moreover, the (NH4)2SO4 supplement enhanced the lipid productivity and gave the maximum yields of biomass and lipid of 15.2 g/L and 9.15 g/L (60.2%), respectively. The C16 and C18 fatty acids were found as main components included oleic acid (55.8%), palmitic acid (16.6%), linoleic acid (11.9%), and stearic acid (7.8%). These results present the possibility to convert the sugars in pineapple pulp hydrolysate to lipids. The fatty acid profile was also similar to vegetable oils. Thus, it could be used as potential feedstock for biodiesel production.

Identification of bioactive peptide from Oreochromis niloticus skin gelatin.

Fish skin, one type of wastes generated from Nile tilapia processing, is still a good source of collagen and gelatin. Bioactive peptides can be obtained from Nile tilapia skin gelatin by trypsin digestion. Trypsin hydrolysate was subsequently purified by gel filtration chromatography. Trypsin A fraction showed the greatest reducing power (5.138 ± 1.060 µM trolox/mg peptide) among all hydrolysate fractions, while trypsin B fraction from gel filtration column was found to exhibit the best radical scavenging and angiotensin-I-converting enzyme (ACE) inhibitory activities 8.16 ± 2.18 µg trolox/mg peptide and 59.32 ± 9.97 % inhibition, respectively. The most active fraction was subjected to MALDI-TOF/TOF MS/MS. After annotation by Mascot sequence matching software (Matrix Science) with Ludwig NR Database, two peptide sequences were identified; GPEGPAGAR (MW 810.87 Da) and GETGPAGPAGAAGPAGPR (MW 1490.61 Da). The docking analysis suggested that the shape of the shorter peptide may be slightly more proper, to fit into the binding cleft of the ACE. However, the binding affinities calculated from the docking showed no significant difference between the two peptides. In good agreement with the in silico data, results from the in vitro ACE inhibitory activity with synthetic peptides also showed no significant difference. Both peptides are thus interesting novel candidates suitable for further development as ACE inhibitory and antioxidant agents from the natural source.

Antioxidant and antihypertensive activity of gelatin hydrolysate from Nile tilapia skin.

Fish skin, a by-product from fish processing industries, still contains a significant amount of protein-rich material. Gelatin was extracted from Nile tilapia skin with the yield 20.77 ± 0.80 % wet weight. Gelatin was then separately hydrolyzed by proteases, including bromelain, papain, trypsin, flavourzyme, alcalase and neutrase. Low molecular weight gelatin hydrolysate (<10 kDa) has a great potential as an antioxidant agent. Flavourzyme hydrolysate has potent activity on ABTS radical scavenging (1,413.61 ± 88.74 µg trolox/mg protein) and also inhibits the oxidation of linoleic acid at a high level (59.74 ± 16.57 % inhibition). The greatest reducing power is in alcalase hydrolysate (4.951 ± 1.577 mM trolox/mg protein). While, bromelain hydrolysate has the highest ferrous ion chelating activity (86.895 ± 0.061 %). Evaluation of the angiotensin-I-converting enzyme's inhibitory activity indicates that all hydrolysates have great potency as an antihypertensive agent. All studied tilapia skin gelatin hydrolysates contain potent antioxidant and anti-hypertensive effects.

Identification of major phenolic compounds from Nephelium lappaceum L. and their antioxidant activities.

Nephelium lappaceum is a tropical fruit whose peel possesses antioxidant properties. Experiments on the isolation and identification of the active constituents were conducted, and on their antioxidant activity using a lipid peroxidation inhibition assay. The methanolic extract of N. lappaceum peels exhibited strong antioxidant properties. Sephadex LH-20 chromatography was utilized in the isolation of each constituent and the antioxidant properties of each was studied. The isolated compounds were identified as ellagic acid (EA) (1), corilagin (2) and geraniin (3). These compounds accounted for 69.3% of methanolic extract, with geraniin (56.8%) as the major component, and exhibited much greater antioxidant activities than BHT in both lipid peroxidation (77-186 fold) and DPPH* (42-87 fold) assays. The results suggest that the isolated ellagitannins, as the principal components of rambutan peels, could be further utilized as both a medicine and in the food industry.

Improvement of myrosinase activity of Aspergillus sp. NR4617 by chemical mutagenesis

A myrosinase (thioglucoside glucohydrolase or thioglucosidase, EC 3.2.3.147) producing fungus, Aspergillus sp. NR4617, was newly isolated from decayed soil sample obtained in Thailand and was subjected to single exposure to two chemical mutagens, ethyl methanesulfonate (EMS) and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Its myrosinase production was selected on low cost medium prepared from mustard seed cake (Brassica juncea). Studies of production and stability of the enzyme showed that EMS mutagenesis increased myrosinase activity. Aspergillus sp. NR4617E1 produced myrosinase 1.90 U ml-1 at 36 hrs of the cultivation equivalent to 171 percent of the enzyme production in wild-type. The stability studies revealed that myrosinase from the mutant strains retained activity similar to wild-type at 30ºC. Aspergillus sp. NR4617E1 degraded 10 mM of glucosinolate completely in 36 hrs. Enhanced myrosinase production and high yields of products (allylisothiocyanate) demonstrated that this mutant could be a new found candidate for feed detoxification and industrial allylisothiocyanate production.

Screening of filamentous fungi for production of myrosinase

A linhagem Aspergillus sp. NR46FB, isolada de solo através da técnica do ágar sulfato de bário-sinigrina, foi testada quanto à producão de mirosinase. O fungo degradou completamente o glicosinolato e produziu 3,19 U.mL-1 de mirosinase, após 48 h de cultivo. Devido à alta producão de mirosinase, esse novo isolado é um potente candidato para aplicacões industriais.

Biodegradation of glucosinolates in brown mustard seed meal (Brassica juncea) by Aspergillus sp. NR-4201 in liquid and solid-state cultures.

Aspergillus sp. NR-4201 was assessed by degrading glucosinolates in brown mustard seed meal (Brassica juncea). A liquid culture of the strain, in a medium derived from the meal, produced total degradation of glucosinolates at 32 h. Under these conditions, the glucosinolate-breakdown product, allylcyanide, was formed in culture filtrates. In a plate culture under sterile conditions, the growth of the strain in heat-treated meal media was shown to be effective at 30 degrees C with 51% moisture, as determined by the measurement of the colony growth rate. On the laboratory scale, solid-state culture under the same conditions gave rise to total glucosinolate degradation within 48 h. In comparison, under non-sterile conditions in either heat-treated or non heat-treated meal samples, the degradations were complete after 60 and 96 h, respectively. In these cases, growth was associated with some out-growths of contaminating fungi, mainly Rhizopus sp. and Mucor sp. The glucosinolate-breakdown product, allylcyanide, was not detected in the solid-state meal-media culture presumably due to evaporative loss from the fermentation matrix.