Optimization of 5-CQA Extraction Conditions from Green Coffee By-Product (Coffea arabica) Using a Response-Surface Design and the Study of Its Extraction Kinetics.

To take advantage of the residues generated in the production of products from green coffee and due to the special interest in the compounds contained in the bean, a by-product obtained after the extraction of the oil was studied. The physical characterization of the green-coffee-bean by-product was carried out. Subsequently, the extraction of compound 5-CQA was carried out via leaching using central composition design 24 and evaluating factors such as temperature, time, solid/solvent ratio, and ethanol percentage, and its yield was quantified using HPLC. In addition, the response-surface methodology was used to maximize the efficiency of 5-CQA extraction and to perform the kinetic study. Yields of 59 ± 2 mg of 5-CQA/g from the by-product were obtained, and by selecting the best leaching conditions, the kinetic study was performed at 45, 60, and 75 °C, increasing the yield to a total of 61.8 ± 3 mg of 5-CQA/g. By applying the kinetic model of mass transfer, a fit of R2 > 0.97 was obtained, with KLa values between 0.266 and 0.320 min−1. This study showed an approach to optimize the 5-CQA extraction conditions, resulting in a simple, fast, reproducible, accurate, and low-cost method.

Succinic acid production with Actinobacillus succinogenes ZT-130 in the presence of succinic acid.

Glucose fermentation with Actinobacillus succinogenes was carried out at different initial concentrations of succinic acid (SA(0)) to determine its effect on growth and on the production of succinic acid itself. The specific rates of biomass production, succinic, formic and acetic acids decreased with SA(0) (0-40 g/l). The partially dissociated form of succinic acid had a higher effect on cell growth and production of succinic acid as compared to the non-dissociated forms of the acids, a fact that has not been reported until now. Cell growth fitted the Jerusalimski model, and the Aiba-Shoda model was suitable for quantification of the inhibition for the production of succinic acid. The growth inhibition constants K(is) and K(ip) and their summatory were consistent with the experimental values obtained, i.e., 22 g/l for the produced acids and 38 g/l for total acids that were the limits at which the biomass formation ceased.

Anaerobic treatment of Tequila vinasses in a CSTR-type digester.

Tequila industries in general produce great volumes of effluents with high pollutant loads, which are discharged (untreated or partially treated) into natural receivers, thus causing severe environmental problems. In this contribution, we propose an integrated system as a first step to comply with the Mexican ecological norms and stabilize the anaerobic treatment of Tequila vinasses with main design criteria: simple and easy operation, reduce operating time and associated costs (maintenance), integrated and compact design, minimal cost of set-up, start-up, monitoring and control. This system is composed of a fully instrumented and automated lab-scale CSTR-type digester, on-line measuring devices of key variables (pH, temperature, flow rates, etc.), which are used along with off-line readings of chemical oxygen demand (COD), biogas composition, alkalinity and volatile fatty acids to guarantee the operational stability of the anaerobic digestion process. The system performance was evaluated for 200 days and the experimental results show that even under the influence of load disturbances, it is possible to reduce the COD concentration to 85% in the start-up phase and up to 95% during the normal operation phase while producing a biogas with a methane composition greater than 65%. It is also shown that in order to maintain an efficient treatment, the buffering capacity (given by the alkalinity ratio, alpha = intermediate alkalinity/total alkalinity) must be closely monitored.

Multiple response optimization analysis for pretreatments of Tequila's stillages for VFAs and hydrogen production.

The objective of this work was study the effect of three pretreatments (alkalinization, thermical treatment, and sonication) on Tequila's stillages hydrolysis process in acidogenesis stage, through the following response variables: soluble chemical oxygen demand (CODs), total sugar and volatile fatty acids profile and the hydrogen production at the time. The stillages were subject to these pretreatments (according to a 2(3) factorial design); afterward they were transferred to a batch reactor at 35 degrees C and inoculated with an anaerobic digestor sludge. Multiple response optimization (MRO) analysis was done to find the global optimum for the response variables described above. This optimum is able to maximize simultaneously all these variables. It was found adequate to be useful hydrolyzing the organic matter present in Tequila's stillages. Mathematical models were fitted to observe the estimated effects of pretreatments on each response variable, then the MRO was applied.

Bagasse hydrolyzates from Agave tequilana as substrates for succinic acid production by Actinobacillus succinogenes in batch and repeated batch reactor.

The aim of this work was to obtain fermentable sugars by enzymatic or acid hydrolyses of Agave tequilana Weber bagasse in order to produce succinic acid with Actinobacillus succinogenes. Hydrolyses were carried out with mineral acids (sulfuric and hydrochloric acids) or a commercial cellulolytic enzyme, and were optimized statistically by a response surface methodology, having as factors the concentration of acid/enzyme and time of hydrolysis. The concentration of sugars obtained at optimal conditions for each hydrolysis were 21.7, 22.4y 19.8g/L for H2SO4, HCl and the enzymatic preparation respectively. Concerning succinic acid production, the enzymatic hydrolyzates resulted in the highest yield (0.446g/g) and productivity (0.57g/Lh) using A. succinogenes in a batch reactor system. Repeated batch fermentation with immobilized A. succinogenes in agar and with the enzymatic hydrolyzates resulted in a maximum concentration of succinic acid of 33.6g/L from 87.2g/L monosaccharides after 5 cycles in 40h, obtaining a productivity of 1.32g/Lh.

Immobilization of Actinobacillus succinogenes by adhesion or entrapment for the production of succinic acid.

The production of succinic acid was studied with entrapped and adsorbed Actinobacillus succinogenes. The adsorption of fermentation products (organic acids in the concentration range of 1-20 g/L) on different supports was evaluated. It was found that succinic acid was adsorbed in small quantities on diatomite and zeolite (12.6 mg/g support). The highest production of succinic acid was achieved with A. succinogenes entrapped in agar beads. Batch fermentations with immobilized cells were carried out with glucose concentrations ranging from 20 to 80 g/L. Succinic acid (43.4 g/L) was obtained from 78.3g/L glucose, and a high productivity (2.83 g/Lh) was obtained with a glucose concentration of 37.6g/L. For repeated batch fermentations (5 cycles in 72 h) with immobilized cells in agar, the total glucose consumed was 147.55 g/L, while the production of succinic acid was 107 g/L. Immobilized cells reduced significantly the fermentation time, yield, productivity and final concentration of succinic acid.

Overexpression of ADH1 and HXT1 genes in the yeast Saccharomyces cerevisiae improves the fermentative efficiency during tequila elaboration.

This work assessed the effect of the overexpression of ADH1 and HXT1 genes in the Saccharomyces cerevisiae AR5 strain during fermentation of Agave tequilana Weber blue variety must. Both genes were cloned individually and simultaneously into a yeast centromere plasmid. Two transformant strains overexpressing ADH1 and HXT1 individually and one strain overexpressing both genes were randomly selected and named A1, A3 and A5 respectively. Overexpression effect on growth and ethanol production of the A1, A3 and A5 strains was evaluated in fermentative conditions in A. tequilana Weber blue variety must and YPD medium. During growth in YPD and Agave media, all the recombinant strains showed lower cell mass formation than the wild type AR5 strain. Adh enzymatic activity in the recombinant strains A1 and A5 cultivated in A. tequilana and YPD medium was higher than in the wild type. The overexpression of both genes individually and simultaneously had no significant effect on ethanol formation; however, the fermentative efficiency of the A5 strain increased from 80.33% to 84.57% and 89.40% to 94.29% in YPD and Agave medium respectively.