Lignin triggers irreversible cellulase loss during pretreated lignocellulosic biomass saccharification

Background

Non-productive binding of enzymes to lignin is thought to impede the saccharification efficiency of pretreated lignocellulosic biomass to fermentable sugars. Due to a lack of suitable analytical techniques that track binding of individual enzymes within complex protein mixtures and the difficulty in distinguishing the contribution of productive (binding to specific glycans) versus non-productive (binding to lignin) binding of cellulases to lignocellulose, there is currently a poor understanding of individual enzyme adsorption to lignin during the time course of pretreated biomass saccharification.

Results

In this study, we have utilized an FPLC (fast protein liquid chromatography)-based methodology to quantify free Trichoderma reesei cellulases (namely CBH I, CBH II, and EG I) concentration within a complex hydrolyzate mixture during the varying time course of biomass saccharification. Three pretreated corn stover (CS) samples were included in this study: Ammonia Fiber Expansion^a (AFEX?-CS), dilute acid (DA-CS), and ionic liquid (IL-CS) pretreatments. The relative fraction of bound individual cellulases varied depending not only on the pretreated biomass type (and lignin abundance) but also on the type of cellulase. Acid pretreated biomass had the highest levels of non-recoverable cellulases, while ionic liquid pretreated biomass had the highest overall cellulase recovery. CBH II has the lowest thermal stability among the three T. reesei cellulases tested. By preparing recombinant family 1 carbohydrate binding module (CBM) fusion proteins, we have shown that family 1 CBMs are highly implicated in the non-productive binding of full-length T. reesei cellulases to lignin.

Conclusions

Our findings aid in further understanding the complex mechanisms of non-productive binding of cellulases to pretreated lignocellulosic biomass. Developing optimized pretreatment processes with reduced or modified lignin content to minimize non-productive enzyme binding or engineering pretreatment-specific, low-lignin binding cellulases will improve enzyme specific activity, facilitate enzyme recycling, and thereby permit production of cheaper biofuels.

     

Prediction of body volume by a stepwise linear regression technique

Body volume and 35 anthropometric measurements were obtained from 88 active soldiers using standard techniques. These anthropometric measurements were examined for their possible relationships to body volume using stepwise linear regression analysis. Four measurements (Body weight, anterior thigh skinfold thickness, subscapular skinfold thickness and suprailiac skinfold thickness) accounted for 99.7% of the variation in body volume and the introduction of each of these measurements in the equation was significant. The regression equation for predicting body volume from these 4 anthropometric measurements had a multiple correlation coefficient of 0.9987 (P less than 0.001). Body weight alone was correlated with body volume to the extent of 0.9966. An attempt has therefore been made to develop a multiple linear regression equation without incorporation of body weight in the regression analysis. Nine measurements were selected by stepwise linear regression analysis for predicting body volume. These nine measurements accounted for 97.1% of the variation in body volume. These equations have been validated on another small sample of 22 soldiers. The analysis has also revealed that a direct regression of body density from the anthropometric variables gives more accurate results than when estimated body volumes are utilized for calculating body density.     

The Maloti minnow Pseudobarbus quathlambae (Barnard, 1938) is not extinct in South Africa

Despite concerted surveys, the Maloti minnow Pseudobarbus quathlambae (Barnard 1938) had not been recorded in South African waters for almost eighty years since the original collections were made at the type locality in the upper uMkhomazana River in 1938. The species was therefore declared extinct in South Africa, whereas extant populations were considered confined to various rivers in the Lesotho highlands. In April 2017, however, this species was rediscovered in the Mzimkhulu River system in KwaZulu-Natal. The rediscovery of a species that was considered locally extinct supports the need for extensive surveys to determine its distribution range, estimate population sizes, assess conservation status and implement effective strategies to ensure its continued existence in KwaZulu-Natal.     

Prediction of GTP interacting residues,dipeptides and tripeptides in a protein from its evolutionary information

Background  

Guanosine triphosphate (GTP)-binding proteins play an important role in regulation of G-protein. Thus prediction of GTP interacting residues in a protein is one of the major challenges in the field of the computational biology. In this study, an attempt has been made to develop a computational method for predicting GTP interacting residues in a protein with high accuracy (Acc), precision (Prec) and recall (Rc).