Журнал биопереработки и биотехники

Biotechnology is a key area of industrial interest and the importance of effective knowledge management for rapid bioprocess development, optimisation and operation is widely recognised as an important driver of biomanufacturing excellence. The Britest suite of tools and methodologies, designed to highlight knowledge gaps within chemical and physical processes, is explored as an approach to bioprocess knowledge acquisition and management. These tools can help identify where optimisation may be most beneficial, and also increase understanding of the process as a whole across a range of disciplines. This research identifies areas where Britest tools are not directly transferable into biotechnological applications, and formulates a whole bioprocess development methodology. The Britest tools have been considered using SuperPro Designer in relation to production of insulin using E. coli. Some of the existing Britest tools have been found to be directly applicable to biological processes, although adaptations were required in some cases, to account for differences between chemical and biological processing. A gap was identified relating to considering the process as a whole, and so a new tool (the Reaction/Reagent/Transformation Tracker, R2T2) was developed to address this. The Britest tools show promise within the context of a bioprocess, although further work is required to fully realise their potential in this exciting field. It is anticipated that the tools can be applied to aid in the identification of Critical Quality Attributes (CQAs) and Critical Process Parameters (CPPs), constructing a robust design space, and facilitating the development of a Quality by Design (QbD) approach to bioprocessing.

Bladder cancer is a heterogeneous disease and occupies highest incidence in developed country. Time-resolved fluoroimmunoassays (TRFIA) is a new detection technique with a feature of sensitive, simple and inexpensive. The aim of this study is to establish a dual-label TRFIA for the simultaneous detection of BLCA-4 and NMP52 in urine in a single run. The sandwich immunoassay was used to detect the concentration of BLCA-4 and NMP52 in urine. BLCA- 4 and NMP52 in urine were captured by anti-BLCA-4 and anti-NMP52 antibodies immobilized on microtiter wells, and then banded together with another anti-BLCA-4 and anti-NMP52 labeled with europium(III) Sm3+ and samarium(III) Eu3+ chelates, followed by fluorescence measurement using time resolved fluorometry. To assess the performance of this assay, clinical urine samples were used, and then commercialized kits were compared. The sensitivity for BLCA-4 and NMP52 detection of this assay were 2 U/mL (dynamic range, 5-300 U/L) and 1 μg/ml (dynamic range, 2-150 μg/ml) respectively. The correlation coefficients (R) between the present dual-label TRFIA and commercially available kits were high. R-values were 0.99 for BLCA-4 and NMP52. The cross-reactivity seems not to influence the results. The present dual-label TRFIA, allowing the simultaneous detection ofBLCA-4 and NMP52, has high sensitivity, specificity, and accuracy in clinical sample analysis. Therefore, it has good prospects of application.

Insects are the most successful animal on earth; the gut microbes present might play an important role in food digestion as well as the interaction with hosts. In this study insect Gryllotalpa krishnani has been used to isolate the symbiotic associated organisms which play role in host metabolism, promote efficient digestion and to protect the host from the harmful microbes. In the present study, microorganisms were isolated from the gut of G. krishnani and characterized for tannase enzyme activity. It was observed that 5 isolates (TAH 6, TAH 13, TAH 36, TAH 38 and TAH 41) out of total 120 tested were able to show significant growth on tannic acid plate. Among these five potential isolates, strain TAH 41 exhibited maximum tannase activity in tannase plate assay and was selected for the further study. The bacterial strain TAH41 was analyzed for biochemical analysis, 16S rDNA sequencing and the result confirmed the strain as Enterobacter cloacae. HPLC analysis showed the formation two peaks representing gallic acid and glucose as a by-product FT-IR analysis also confirmed the same. The polyacrylamide gel electrophoresis (SDS-PAGE) and zymogram analysis showed the molecular mass of tannase enzyme in cell free extract was ~45 kDa, the analysis suggested this tannase enzyme to be one of the smallest of the bacterial source, which could be attributed to the formation of di or tri‑galloyl glucose. The present study was the first report E. cloacae with tannase activity from G. krishnani gut. E. cloacae which may endow the insect with some ecological advantages by enabling them to overcome the anti-nutritional effects of plant tannins.