Biotechnological potential of industrial wastes for economical citric acid bioproduction by Aspergillus niger through submerged fermentation

Submerged citric acid (CA) bioproduction was carried out by Aspergillus niger NRRL‐567 using various industrial wastes, such as brewery spent liquid (BSL), lactoserum and starch industry water sludge. CA bioproduction was carried out by varying the temperature (25–35 °C), pH (3–5), addition of inducers, incubation time and supplementation with different proportions of apple pomace ultrafiltration sludge (APS). The results indicated that under the best conditions with 3% (v/v) methanol, the optimal concentration of 11.34 g L^?1 CA was recorded using BSL at pH 3.5 and temperature 30 °C after 120‐h incubation period. Supplementation of methanol resulted in an increase of 56% CA production. Meanwhile, under similar conditions, higher concentration of 18.34 g L^?1 CA was reported with the supplementation of BSL with 40% (v/v) APS having suspended solids concentration of 30 g L^?1. The present study demonstrated the potential of BSL supplemented with APS as an alternative cheap substrate for CA fermentation.     

Construction of the mutant strain in Aspergillus oryzae 3.042 for abundant proteinase production by the N+ ion implantation mutagenesis

Because of secreting large amounts of enzymes, Aspergillus oryzae was widely used in the fermentation of soy sauce in many Asian countries. Here, N⁺ ion implantation mutagenesis was applied to improve the ability of A. oryzae to secrete acid protease. Several mutants were screened using three kinds of plates (Casein medium, Czapek’s medium and carboxymethyl cellulose medium agar plates). Compared with the other mutants, the mutant A100‐8 could secrete the most protease. Acid protease activity of A100‐8 was enhanced about 44.1% at 36 h by koji fermentation test. In addition, A100‐8 was stable by periodically subculturing and maintaining on the agar slant. The mRNA expression levels of two kinds of acid protease (serine carboxypeptidase and aspartyl protease) were measured by RT‐PCR at different periods. Serine carboxypeptidase and some kinds of aspartyl protease of A100‐8 were significantly (P < 0.01) expressed higher than the control at all times.     

Biochemical characterisation and application of lipases produced by Aspergillus sp. on solid‐state fermentation using three substrates

Lipase from Aspergillus sp. obtained by solid‐state fermentation (SSF) on wheat bran (LWB), soybean bran (LSB) and soybean bran combined with sugarcane bagasse (LSBBC) were 67.5, 58 and 57.3 U of crude lipase per gram substrate, respectively. The optimum pH of activity and stability of the LWB was between 8 and 9, and the optimum temperature of activity and stability was 50 °C and up to 60 °C, respectively. The LSB and LSBBC showed two peaks of optimum pH (4 and 6) and optimal values of temperature and stability at 50 °C. The LSB was stable in the pH range of 6–7, while LSBBC in the range of pH 4–7. All the enzymes show activities on p‐nitrophenyl esters (butyrate, laurate and palmitate). LWB stood out either on the hydrolysis of sunflower oil, presenting 66.1% of the activity over commercial lipase and on the esterification of oleic acid and ethanol, surpassing the activities of the commercial lipases studied. The thin layer chromatography showed that LWB and LSB have produced ethyl esters from corn oil, while LWB produced it from sunflower oil.     

Effect of Capsicum carotenoids on growth and aflatoxins production by Aspergillus flavus isolated from paprika and chilli

The aim of this study was to determine the effect of a carotenoid mixture (Capsantal FS-30-NT), containing capsanthin and capsorubin, on growth and aflatoxins (AF) production of AF-producing Aspergillus flavus isolates.     

An Iterative O‐Methyltransferase Catalyzes 1,11‐Dimethylation of Aspergillus fumigatus Fumaric Acid Amides

S‐adenosyl‐l ‐methionine (SAM)‐dependent methyltransfer is a common biosynthetic strategy to modify natural products. We investigated the previously uncharacterized Aspergillus fumigatus methyltransferase FtpM, which is encoded next to the bimodular fumaric acid amide synthetase FtpA. Structure elucidation of two new A. fumigatus natural products, the 1,11‐dimethyl esters of fumaryl‐l ‐tyrosine and fumaryl‐l ‐phenylalanine, together with ftpM gene disruption suggested that FtpM catalyzes iterative methylation. Final evidence that a single enzyme repeatedly acts on fumaric acid amides came from an in vitro biochemical investigation with recombinantly produced FtpM. Size‐exclusion chromatography indicated that this methyltransferase is active as a dimer. As ftpA and ftpM homologues are found clustered in other fungi, we expect our work will help to identify and annotate natural product biosynthesis genes in various species.     

Production and analysis of ochratoxin A produced by Aspergillus ochraceus ITEM 5137 in submerged culture

A rapid simple and economical method was described for the determination of ochratoxin A produced by Aspergillusochraceus ITEM 5117 grown in a biofermenter in submerged culture. The ochratoxin A was determinate with RP-HPLC-FLD (reverse phase high performance liquid chromatography) with direct injection in the HPLC apparatus using a C18 column. The mycotoxin was completely resolved by using the mixture acetonitrile, water and acetic acid (49:49:2 v/v) as the mobile phase with a flow rate of 1.0 mL/min. Mean recoveries of ochratoxin A ranged from 95.36% to 103.15%. The limit of detection for ochratoxin A in medium was found to be 1 μg L⁻¹.     

Delivery systems for biological control agents to manage aflatoxin contamination of pre-harvest maize

While soil application of a competitive non-toxigenic Aspergillus flavus strains is successful in reducing aflatoxin contamination in certain crops, direct application to aerial reproductive structures could be more effective for maize. A sprayable, clay-based water-dispersible granule formulation was developed to deliver non-toxigenic A. flavus strain K49 directly to maize ears. The efficacy of the K49 water-dispersible granule in mitigating aflatoxin in maize (Zea mays L.) was evaluated. Field studies were conducted to compare K49 colonization and effectiveness in reducing aflatoxin contamination when applied either as a soil inoculant or as a directed spray in plots infested with toxigenic strain F3W4. Fifty percent of non-toxigenic A. flavus was recovered from non-treated controls and from plots soil inoculated with K49 on wheat. In spray treatments with formulated or unformulated K49 conidia, over 90% of A. flavus recovered was non-toxigenic. Soil-applied K49 reduced aflatoxin contamination by 65% and spray applications reduced contamination by 97%. These findings suggest direct spray application of non-toxigenic A. flavus strains may be better than soil inoculation at controlling maize aflatoxin contamination and that a water-dispersible granule is a viable delivery system for maintaining viability and efficacy of the biological control agent, K49.     

Kinetics and thermodynamics of glucoamylase inhibition by lactate during fermentable sugar production from food waste

BACKGROUND: Glucoamylase hydrolysis is a key step in the bioconversion of food waste with complicated composition. This work investigated the effect of lactate on glucoamylase from Aspergillus niger UV‐60, and inhibition mechanisms of glucoamylase by lactate during food waste hydrolysis. RESULTS: For 125 min hydrolysis of food waste (10%, dry basis), reducing sugars produced in the absence of lactate were 15%, 26% and 56% more than those produced in the presence of 24 g L^?1 lactate at 60, 50 and 40 °C, respectively. Kinetic study showed that the type of glucoamylase inhibition by lactate was competitive, and K_m (Michaelis‐Menten constent), V_max (maximum initial velocity), K_I (inhibition constant) were 103.2 g L^?1, 5.0 g L^?1 min^?1, 100.6 g L^?1, respectively, for food waste hydrolysis at 60 °C and pH 4.6. Lactate also accelerated glucoamylase denaturation significantly. Activation energy of denaturation without inhibitor was 61% greater than that of denaturation with inhibitor (24 g L^?1 lactate). Half‐lives (t_1/2) without inhibitor were 7.6, 2.7, 2.6, 1.7 and 1.2 times longer than those with inhibitor at temperature 40, 45, 50, 55 and 60 °C, respectively. CONCLUSION: These results are helpful to process optimization of saccharification and bioconversion of food waste. Copyright © 2010 Society of Chemical Industry     

Effects of rice koji inoculated with Aspergillus luchuensis on the biochemical and sensory properties of a sailfin sandfish (Arctoscopus japonicus) fish sauce

The effects of rice koji inoculated with Aspergillus luchuensis on the biochemical and sensory properties of fish sauce produced from sailfin sandfish (Arctoscopus japonicus) with different salt concentrations (10% and 20%) were investigated over 10 months. Fish sauces prepared from sailfin sandfish with rice koji and a low salt concentration had higher total nitrogen, amino nitrogen and total free amino acid than did those without rice koji or with a high salt concentration. Sensory evaluation of fish sauces prepared with rice koji and a low salt concentration found better taste, flavour and overall acceptance than those prepared without rice koji during fermentation (P < 0.05). The use of rice koji increased the total nitrogen, amino nitrogen and free amino acid and also enhanced the sensory qualities during fermentation. These results suggest that the use of rice koji can result in a low‐salt fish sauce with better flavour.