Using poly(N-Vinylcaprolactam) to Improve the Enzymatic Hydrolysis Efficiency of Phenylsulfonic Acid-Pretreated Bamboo

Chemical pretreatment followed by enzymatic hydrolysis has been regarded as a viable way to produce fermentable sugars. Phenylsulfonic acid (PSA) pretreatment could efficiently fractionate the non-cellulosic components (hemicelluloses and lignin) from bamboo and result in increased cellulose accessibility that was 10 times that of untreated bamboo. However, deposited lignin could trigger non-productive adsorption to enzymes, which therefore significantly decreased the enzymatic hydrolysis efficiency of PSA-pretreated bamboo substrates. Herein, poly(N-vinylcaprolactam) (PNVCL), a non-ionic surfactant, was developed as a novel additive for overcoming the non-productive adsorption of lignin during enzymatic hydrolysis. PNVCL was found to be not only more effective than those of commonly used lignosulfonate and polyvinyl alcohol for overcoming the negative effect of lignin, but also comparable to the robust Tween 20 and bovine serum albumin additives. A PNVCL loading at 1.2 g/L during enzymatic hydrolysis of PSA pretreated bamboo substrate could achieve an 80% cellulosic enzymatic conversion and meanwhile reduce the cellulase loading by three times as compared to that without additive. Mechanistic investigations indicated that PNVCL could block lignin residues through hydrophobic interactions and the resultant PNVCL coating resisted the adsorption of cellulase via electrostatic repulsion and/or hydration. This practical method can improve the lignocellulosic enzymatic hydrolysis efficiency and thereby increase the productivity and profitability of biorefinery.

Hydrogen peroxide signaling via its transformation to a stereospecific alkyl hydroperoxide that escapes reductive inactivation

During systemic inflammation, indoleamine 2,3-dioxygenase 1 (IDO1) becomes expressed in endothelial cells where it uses hydrogen peroxide (H2O2) to oxidize L-tryptophan to the tricyclic hydroperoxide, cis-WOOH, that then relaxes arteries via oxidation of protein kinase G 1α. Here we show that arterial glutathione peroxidases and peroxiredoxins that rapidly eliminate H2O2, have little impact on relaxation of IDO1-expressing arteries, and that purified IDO1 forms cis-WOOH in the presence of peroxiredoxin 2. cis-WOOH oxidizes protein thiols in a selective and stereospecific manner. Compared with its epimer trans-WOOH and H2O2, cis-WOOH reacts slower with the major arterial forms of glutathione peroxidases and peroxiredoxins while it reacts more readily with its target, protein kinase G 1α. Our results indicate a paradigm of redox signaling by H2O2 via its enzymatic conversion to an amino acid-derived hydroperoxide that ‘escapes’ effective reductive inactivation to engage in selective oxidative activation of key target proteins.

ECMO for Metabolic Crisis in a Patient with Mitochondrial Disease

Patients with mitochondrial disease exhibit disrupted pyruvate oxidation, resulting in intraoperative and perioperative physiologic derangements. Increased enzymatic conversion of pyruvate via lactate dehydrogenase during periods of fasting or stress can lead to metabolic decompensation, with rapid development of fatal lactic acidosis. We describe the intraoperative management and postoperative critical care of a patient with mitochondrial disease who presented for repair of esophageal perforation following repair of a paraesophageal hernia. His surgery was complicated by the development of metabolic crisis and severe lactic acidosis which became resistant to conventional therapy before ultimately resolving with the initiation of venoarterial extracorporeal membrane oxygenation (VA-ECMO).

KINETIC STUDY COMPARISON OF IMMOBILIZED GLUCOSE ISOMERASE (GENSWEET AND SWEETZYME IT) IN STIRRED TANK REACTOR AND PACKED BED REACTOR

D- Glucose/xylose isomerase catalysis the reversible isomerization of aldoses to ketoses such as D-glucose and D-xylose to D-fructose and D-xylose respectively. High fructose corn syrup (HFCS), a low calorie sugar substitute for cane sugar, utilizes Glucose isomerase enzyme for conversion of glucose to fructose. The conversion of glucose to fructose favours more at high temperature, providing an incentive to utilize thermostable and thermoactive glucose isomerase in High fructose corn syrup (HFCS) production. Present studies emphasize on enzymatic conversion and optimization using Sweetzyme IT extra & Gensweet, commercially available glucose isomerases. The experiments were carried out for enzymatic conversion of glucose to fructose using Gensweet and Sweetzyme in Packed bed reactor (PBR) and Stirred tank reactor (STR). Maximum conversion was seen in Stirred tank reactor (STR) using both of these enzymes, approx 10 % more Fructose conversion comparing it to packed bed reactor (PBR). Also, Stirred tank reactor (STR) reaction conditions such as pH, buffers, cofactor (MgSO4) requirement were optimized to achieve optimum enzyme activity. Analysis of enzymatic conversion samples was done using HPLC-RID (using Zorbax Column). The importance of the divalent cation MgCl2 for optimal enzyme activity was investigated. The enzyme performed best at pH 7.5 and 60°C, using 10mM MgSO4 as a cofactor. Utilizing Gensweet in Stirred tank reactor (STR), the maximum fructose transformation was 44 %. The most activity was detected with Sodium phosphate buffers, and EPPS buffers at pH 7 and 8, accordingly, whereas the least activity was reported with TRIS HCl buffer.