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A xyloglucan-specific endo-1,4β-glucanase (XcXGHA) from Xanthomonas citri pv. mangiferaeindicae has been cloned, expressed in Escherichia coli, purified and characterised. The XcXGHA enzyme belongs to CAZy family GH74 and has catalytic site residues conserved with other xyloglucanases in this family. At its optimal reaction conditions, pH 7.0 and 40 °C, the enzyme has a k/ K value of 2.2 × 10 min M on a tamarind seed xyloglucan substrate. XcXGHA is relatively stable within a broad pH range (pH 4-9) and up to 50 °C ( t of 74 min). XcXGHA is proven to be xyloglucan-specific, and a glycan microarray study verifies that XcXGHA catalyses cleavage of xyloglucan extracted from both monocot and dicot plant species. The enzyme catalyses hydrolysis of tamarind xyloglucan in a unique way by cleaving XXXG into XX and XG (X is xylosyl-substituted glucose; G is unsubstituted glucose), is able to degrade more complex xyloglucans and notably is able to cleave near more substituted xyloglucan motifs such as L [i.e. α- l-Fuc p-(1 → 2)-β- d-Gal p-(1 → 2)-α- d-Xyl p-(1 → 6)-β- d-Glc p]. LC-MS/MS analysis of product profiles of tamarind xyloglucan which had been catalytically degraded by XcXGHA revealed that XcXGHA has specificity for X in subsite −1. The 3D model suggests that XcXGHA consists of two seven-bladed β-propeller domains with the catalytic center formed by the interface of these two domains, which is conserved in xyloglucanases in the GH74 family. However, the XcXGHA has two amino acids (D264 and R472) that differ from the conserved residues of other GH74 xyloglucanases. These two amino acids were predicted to be located on the opposite side of the active site pocket, facing each other and forming a closing surface above the active site pocket. These two amino acids may contribute to the unique substrate specificity of the XcXGHA enzyme. [ABSTRACT FROM AUTHOR]

Abstract: In steam pretreatment the defibration is usually achieved by an explosion at the end of the treatment, but can also be carried out in a subsequent refiner step. A steam explosion and a steam refining unit were compared by using the same raw material and pretreatment conditions, i.e. temperature and time. Smaller particle size was needed for the steam explosion unit to obtain homogenous slurries without considerable amounts of solid chips. A higher amount of volatiles could be condensed from the vapour phase after steam refining. The results from enzymatic hydrolysis showed no significant differences. It could be shown that, beside the chemical changes in the cell wall, the decrease of the particle size is the decisive factor to enhance the enzymatic accessibility while the explosion effect is not required. [Copyright &y& Elsevier]

Abstract: The conversion of wheat straw to fermentable sugar for bioethanol production typically involves a thermal pretreatment step, followed by enzymatic hydrolysis. In this study we have investigated the effect of steam explosion parameters on wheat straw digestibility using a newly designed steam explosion unit and a process without acid impregnation. The wheat straw was steam pretreated using 18 different conditions in the temperature range of 170–220 °C and the resulting material was used directly (i.e. without washing) for enzymatic hydrolysis and fermentation in either a separate hydrolysis and fermentation (SHF)-type or a simultaneous saccharification and fermentation (SSF)-type set-up. Maximum glucose yields upon enzymatic hydrolysis were obtained after pretreatment at 210 °C for 10 min and yields were similar at harsher conditions. Xylose yields increased with temperature and residence time up to 190 °C, but decreased at harsher pretreatment conditions since these led to xylan degradation and concomitant production of furfural. In an SHF-type set-up ethanol formation did not follow enzymatic glucose release and was inversely correlated with furfural levels. An SFF-type set-up displayed a straightforward correlation between the expected amount of released glucose and the ethanol yields. The highest saccharification yields corresponded to about 90% of the cellulose in the substrate. Overall, this study shows that steam explosion without an acid catalyst is a good pretreatment method for saccharification of wheat straw. Optimal steam explosion conditions need to be a compromise between sugar accessibility and sugar degradation. [Copyright &y& Elsevier]

Solid residues such as primary sludge (PS), waste activated sludge (WAS), and food waste (FW) can be stabilized through anaerobic digestion (AD). Application of the thermal hydrolysis process (THP) prior to AD results in several benefits in AD and dewatering. However, soluble recalcitrant compounds associated with Maillard reactions have been identified after THP which can impact downstream processes and water discharge limits. In this study, the soluble colloidal chemical oxygen demand, color, ultraviolet absorbance at 254 nm and dissolved organic nitrogen in seven full‐scale THP facilities were quantified and compared. The THP substrate influenced the concentration of soluble melanoidin‐associated compounds in the digestates. THP implementation in five water resource recovery facilities (WRRFs) was modeled and found to give a 3–8 mg/L increase on the water effluent COD concentration depending on the PS/WAS ratio. The results provide novel information useful in planning new WRRFs and optimization of existing facilities. Practitioner points: High amounts of WAS in substrate resulted in higher concentrations of CODsc, color and dissolved organic nitrogen in the digestate.Food waste treated at 145°C showed equal or lower concentrations of all components compared with sludge operated at 165°C.Installation of THP will increase the COD concentration in the water effluent of a water resource recovery facility.The characteristics of the THP substrate are important to consider in cost/benefit analysis when planning the installation of THP. [ABSTRACT FROM AUTHOR]