a b s t r a c tIn this study, we investigated the enzymatic hydrolysis of pretreated sugarcane bagasse using eight different enzymatic blends obtained from concentrated crude enzyme extracts produced by Penicillium funiculosum and Trichoderma harzianum as well as from the extracts in combination with a commercial enzymatic cocktail. The influence of different levels of biomass delignification, degree of crystallinity of lignicellulose, composition of enzymatic activities and BSA on enzymatic hydrolysis yields (HYs) was evaluated. Our X-ray diffraction studies showed that crystallinity of lignocellulose is not a key determinant of its recalcitrance toward enzymatic hydrolysis. In fact, under the experimental conditions of our study, an increase in crystallinity of lignocellulosic samples resulted in increased glucose release by enzymatic hydrolysis. Furthermore, under the same conditions, the addition of BSA had no significant effect on enzymatic hydrolysis. The most efficient enzyme blends were obtained by mixing a commercial enzymatic cocktail with P. funiculosum or T. harzianum cellulase preparations (HYs above 97%) followed by the concentrated extract of P. funiculosum alone (HY = 88.5%). Increased hydrolytic efficiencies appeared to correlate with having an adequate level of both ␤-glucosidase and xylanase activities in the blends.

This study demonstrates the production of an active enzyme cocktail produced by growing Trichoderma harzianum on sugarcane bagasse. The component enzymes were identified by LCMS-MS. Glycosyl hydrolases were the most abundant class of proteins, representing 67% of total secreted protein. Other carbohydrate active enzymes involved in cell wall deconstruction included lytic polysaccharide mono-oxygenases (AA9), carbohydrate-binding modules, carbohydrate esterases and swollenin, all present at levels of 1%. In total, proteases and lipases represented 5 and 1% of the total secretome, respectively, with the rest of the secretome being made up of proteins of unknown or putative function. This enzyme cocktail was efficient in catalysing the hydrolysis of sugarcane bagasse cellulolignin to fermentable sugars for potential use in ethanol production. Apart from mapping the secretome of T. harzianum, which is a very important tool to understand the catalytic performance of enzyme cocktails, the gene coding for T. harzianum swollenin was expressed in Aspergillus niger. This novel aspect in this work, allowed increasing the swollenin concentration by 95 fold. This is the first report about the heterologous expression of swollenin from T. harzianum, and the findings are of interest in enriching enzyme cocktail with this important accessory protein which takes part in the cellulose amorphogenesis. Despite lacking detectable glycoside activity, the addition of swollenin of T. harzianum increased by two-fold the hydrolysis efficiency of a commercial cellulase cocktail. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:327-336, 2016.

Biosurfactants, a wide group of compounds produced by different microorganisms, generally have less toxicity and are more biodegradable than synthetic surfactants. Biosurfactant-producing bacteria can be found in contaminated environments, such as soils receiving pesticide applications constantly, or in pesticides treatment systems where microorganisms are adapted to biodegrading pesticides. Five pesticide-tolerant bacteria previously isolated from a pesticide biopurification system were evaluated as biosurfactant-producers. Pseudomonas rhodesiae C4, Rhodococcus jialingiae C8 and Pseudomonas marginalis C9 strains were positive in qualitative tests. Biosurfactant production by these strains using Bushnell-Haas medium with olive oil at 2% (w/v) was evaluated as emulsification index, oil displacement, droplet collapse test and surface tension. After 144 h, these strains showed a similar emulsification index of >55%. The two Pseudomonas (C4 and C9) strains showed lower superficial tension compared with Rhodococcus strain (C8)—34.47, 37.44 and 47.55 mN/m for strains C4, C9 and C8, respectively. The chemical characterization of the biosurfactants revealed the presence of glycolipids in P. rhodesiae (C4) and glycopeptides in P. marginalis (C9). The degradation of chlorpyrifos increased from 39.2% to 51.6% when biosurfactants produced by P.rhodesiae (C4) were added (10%) with respect to the control. Therefore, biopurification systems are a relevant source of biosurfactant-producing bacteria with environmental biotechnology applications.

Rhamnolipids (RMLs) have more effectiveness for specific uses according to their homologue proportions. Thus, the novelty of this work was to compare mono‐RMLs and di‐RMLs physicochemical properties on microbial enhanced oil recovery (MEOR) applications. For this, RML produced by three strains of Pseudomonas aeruginosa containing different homologues proportion were used: a mainly mono‐RMLs producer (mono‐RMLs); a mainly di‐RMLs producer (di‐RMLs), and the other one that produces relatively balanced amounts of mono‐RML and di‐RML homologues (mono/di‐RML). For mono‐RML, the most abundant molecules were Rha‐C10C10 (m/z 503.3), for di‐RML were RhaRha‐C10C10 (m/z 649.4) and for Mono/di‐RML were Rha‐C10C10 (m/z 503.3) and RhaRha‐C10C10 (m/z 649.4). All RMLs types presented robustness under high temperature and variation of salinity and pH, and high ability for oil displacement, foam stability, wettability reversal and were classified as safe for environment according to the European Union Directive No. 67/548/EEC. For all these properties, it was observed a highlight for mono‐RML. Mono‐RML presented the lowest surface tension (26.40 mN/m), interfacial tension (1.14 mN/m), and critical micellar concentration (CMC 27.04 mg/L), the highest emulsification index (EI24 100%) and the best wettability reversal (100% with 25 ppm). In addition, mono‐RML showed the best acute toxicity value (454 mg/L), making its application potential even more attractive. Based on the results, it was concluded that all RMLs homologues studied have potential for MEOR applications. However, results showed that mono‐RML stood out and have the best mechanism of oil incorporation in micelles due their most effective surface‐active physicochemical features.

Propionic acid (PA) is an important organic compound with extensive application in different industrial sectors and is currently produced by petrochemical processes. The production of PA by large-scale fermentation processes presents a bottleneck, particularly due to low volumetric productivity. In this context, the present work aimed to produce PA by a biochemical route from a hemicellulosic hydrolysate of sorghum bagasse using the strain Propionibacterium acidipropionici CIP 53164. Conditions were optimized to increase volumetric productivity and process efficiency. Initially, in simple batch fermentation, a final concentration of PA of 17.5 gL −1 was obtained. Next, fed batch operation with free cells was adopted to minimize substrate inhibition. Although a higher concentration of PA was achieved (38.0 gL −1 ), the response variables (Y P/S = 0.409 gg −1 and Q P = 0.198 gL −1 H −1 ) were close to those of the simple batch experiment. Finally, the fermentability of the hemicellulosic hydrolysate was investigated in a sequential batch with immobilized cells. The PA concentration achieved a maximum of 35.3 gL −1 in the third cycle; moreover, the volumetric productivity was almost sixfold higher (1.17 gL −1 H −1 ) in sequential batch than in simple batch fermentation. The results are highly promising, providing preliminary data for studies on scaling up the production of this organic acid.