Production and characterization of alkaline extracellular lipase from newly isolated strain Aspergillus awamori HB-03

A strain HB-03 to produce alkaline extracellular lipase was isolated from oil-rich soil samples and identified as Aspergillus awamori. The growth conditions and nutritional factors for lipase production by strain HB-03 were optimized, and the maximum lipase production of （45.9±2.3） U/mL was obtained at 30 ℃ and pH 7.0 after 36 h using olive oil （1%） and sucrose （0.5%） as carbon sources and combination of peptone （2%）, yeast extract （0.5%） and ammonium sulfate （0.1%） as nitrogen sources. The lipase was purified to homogeneity with 10.6-fold, 18.84% yield and a specific activity of 1 862.2 U/mg using ammonium sulfate precipitation followed by SephadexG-75 gel filtration chromatography. The purified lipase with molecular mass of 68 ku was estimated by SDS-PAGE. The optimum pH and temperature for the purified lipase were found to be 8.5 and 40 ℃, respectively. The lipase kept more than 80% of activity in pH 7.0-10.0 and temperatures up to 45 ℃. The metal ions of Mn2＋, Ba2＋ significantly enhanced the lipase activity, whereas Cu2＋, Fe3＋ and Mg2＋ strongly reduced the lipase activity. The Km and Vmax values of the purified enzyme for p-nitrophenyl palmitate were 0.13 mrnol/L and 60.6 mmol/（L.min）, respectively. The results show that this novel lipase has potential industrial applications.

Water Retention Value： A Study Model-based by Asperglillus awamori and Aspergillus oryzae Embrace Three Models of Solid Substrate

The goal of this study was to evaluate the water retention value （WRV） of a test solid substrate and a fungal cell in solid state fermentation （SSF）. WRV is the ratio of the weight of water retained after centrifugation under specific conditions by a wet sample to the oven dry weight of the same sample. SSF refers to the microbial fermentation, which takes place in the absence or near absence of free water, thus being close to the natural environment. Many factors are involved in a successful SSF process. In addition to biological parameters, the SSF process is also dependent on physical factors such as WRV. A centrifugal technique has been modified and applied to the evaluation of WRV. Wheat bran, soybean hulls and rapeseed meal were used as model substrate. Aspergillus awamori and Aspergillus oryzae were used as model microorganism. Results revealed that the ability of wheat bran to retain water in the solid substrate is 56% higher than that of soybean hulls and rapeseed meal. In the term of fungal cell, the ability of A. oryzae to retain water in the cells was higher （73% higher） than that ofA. awamori. In addition, through oven method moisture content loss from A. awamori is 46% higher than that from A. oryzae during drying process. Nevertheless, it can be seen that A. oryzae is able to retain water content about 5 times higher than A. awamori. Through this results, we found that WRV varies depending on solid substrates and microorganisms. This initial information can be beneficial in the SSF process to be carried out.

Multi-enzymes Production Studies in Single Tray Solid State Fermentation with Opened and Closed System

The robustness ofA. awamori and A. oryzae as enzyme producers is exploited in fungal fermentation on agricultural solid waste. High-level production of extracellular glucoamylase, protease, cellulase and xylanase has been achieved. Three different types of ＇waste＇ solids （wheat bran, soybean hulls and rapeseed meal） have been used in studies of solid state fermentation （SSF）. The enzymes could be produced in significant levels by continuously supplying oxygen （02） through the tray system known as ＂closed＂ and ＂opened＂ tray systems. A perforated tray system was developed in this study that permits direct access to 02. Testing the tray system with different perforated mesh aperture sizes in this study did not yield different results in growth performance of A. awamori and A. oryzae. A. awamori and A. oryzae can be very versatile in producing various enzymes with different substrates with different starch, protein, hemiceilulose and cellulose contents. These studies indicate that A. awamori is more suitable for the efficient production of multiple enzymes in the closed system including xylanase and cellulase, while the production of glucoamylase and protease is superior in the opened system. A. oryzae is more suitable for the efficient production of protease and cellulase in the closed system, while the production of protease is more favourable the opened system. A. awamori efficiently consumed starch in wheat bran medium and produced very high glucoamylase activity, and after that, the fungus efficiently produced other enzymes to degrade other complex nutrients such as protein, hemicellulose and cellulose. Meanwhile, A. oryzae efficiently consumed protein in rapeseed meal and produced very high protease activity. The ability of both filamentous fungi, to convert biomass through SSF bioconversion will have a great impact on food and agro-industry in every aspect of life from food and medicine to fuel.

Selection of Relevant Variables to the Enzyme Production on Red Grape Pomace by Solid-State Fermentation

With the aim of to valorise red grape pomace and to reduce its environmental impact, the production of enzymatic preparations appear as an interesting choice. Statistical experimental Plackett-Burman designs were applied for the selection of relevant medium components and culture conditions for cellulase, xylanase, polygalacturonase and tannase production by Aspergillus awamori, in solid-state fermentation on red grape pomace. Ten variables were tested： initial moisture content （IMC）, particle size （PS）, temperature, initial pH, time of cultivation, mixing （Mx）, and additions of： fructose, tannic acid, sodium phosphate, and ammonium sulphate （ASA）. Results indicate that the production of each enzyme was affected in a distinct way by the different variables. Also, for each of the enzyme activities considered, IMC must be carefully controlled, and optimized above 65%; PS and Mx, must not be taken into account and ASA must be discarded. The other variables studied, must be selected according to the enzyme activity that will be favored.