Consolidated bioprocessing (CBP) of cellulose is a cost-effective route to produce valuable biochemicals by integrating saccharification, fermentation and cellulase synthesis in a single step. However, the lack of understanding of governing factors of interdependent saccharification and fermentation in CBP eludes reliable process optimization. Here, we propose a new framework that synergistically couples population balances (to simulate cellulose depolymerization) and cybernetic models (to model enzymatic regulation of fermentation) to enable improved understanding of CBP. The resulting framework, named the unified cybernetic-population balance model (UC-PBM), enables simulation of CBP driven by coordinated control of enzyme synthesis through closed-loop interactions. UC-PBM considers two key aspects in controlling CBP: (1) heterogeneity in cellulose properties and (2) cellular regulation of competing cell growth and cellulase secretion. In a case study on Clostridium thermocellum, UC-PBM not only provides a decent fit with various exometabolomic data, but also reveals that: (i) growth-decoupled cellulase-secreting pathways are only activated during famine conditions to promote the production of growth substrates, and (ii) starting cellulose concentration has a strong influence on the overall flux distribution. Equipped with mechanisms of cellulose degradation and fermentative regulations, UC-PBM is practical to explore phenotypic functions for primary evaluation of microorganisms' potential for metabolic engineering and optimal design of bioprocess.
A total of 31 strains of Vibrio cholerae O1 (10 from outbreak cases and 7 from surface water) and non-O1 (4 from clinical and 10 from surface water sources) isolated between 1993 and 1997 were examined with respect to presence of cholera enterotoxin (CT) gene by PCR-based assays, resistance to antibiotics, plasmid profiles and random amplified polymorphic DNA (RAPD) analysis. All were resistant to 9 or more of the 17 antibiotics tested. Identical antibiotic resistance patterns of the isolates may indicate that they share a common mode of developing antibiotic resistance. Furthermore, the multiple antibiotic resistance indexing showed that all strains tested originated from high risk contamination. Plasmid profile analysis by agarose gel electrophoresis showed the presence of small plasmids in 12 (7 non-O1 and 5 O1 serotypes) with sizes ranging 1.3-4.6 MDa. The CT gene was detected in all clinical isolates but was present in only 14 (6 O1 serotype and 8 non-O1 serotype) isolates from environmental waters. The genetic relatedness of the clinical and environmental Vibrio cholerae O1 and non-O1 strains was investigated by RAPD fingerprinting with four primers. The four primers generated polymorphisms in all 31 strains of Vibrio cholerae tested, producing bands ranging from < 250 to 4500 bp. The RAPD profiles revealed a wide variability and no correlation with the source of isolation. This study provides evidence that Vibrio cholerae O1 and non-O1 have significant public health implications.
Simultaneous Saccharification and Fermentation (SSF) is a process where microbes have to first excrete extracellular enzymes to break polymeric substrates such as starch or cellulose into edible nutrients, followed by in situ conversion of those nutrients into more valuable metabolites via fermentation. As such, SSF is very attractive as a one-pot synthesis method of biological products. However, due to the co-existence of multiple biochemical steps, modeling SSF faces two major challenges. The first is to capture the successive chain-end and/or random scission of the polymeric substrates over time, which determines the rate of generation of various fermentable substrates. The second is to incorporate the response of microbes, including their preferential substrate utilization, to such a complex broth. Each of the above-mentioned challenges has manifested itself in many related areas, and has been competently but separately attacked with two diametrically different tools, i.e., the Population Balance Modeling (PBM) and the Cybernetic Modeling (CM), respectively. To date, they have yet to be applied in unison on SSF resulting in a general inadequacy or haphazard approaches to examine the dynamics and interactions of depolymerization and fermentation. To overcome this unsatisfactory state of affairs, here, the general linkage between PBM and CM is established to model SSF. A notable feature is the flexible linkage, which allows the individual PBM and CM models to be independently modified to the desired levels of detail. A more general treatment of the secretion of extracellular enzyme is also proposed in the CM model. Through a case study on the growth of a recombinant Saccharomyces cerevisiae capable of excreting a chain-end scission enzyme (glucoamylase) on starch, the interlinked model calibrated using data from the literature (Nakamura et al., Biotechnol. Bioeng. 53:21-25, 1997), captured features not attainable by existing approaches. In particular, the effect of various enzymatic actions on the temporal evolution of the polymer distribution and how the microbes respond to the diverse polymeric environment can be studied through this framework.
The dynamics of droplet breakup during emulsification is a complicated process due to the interplay between multiple physico-chemical and hydrodynamic factors, especially in an energy-intensive ultrasound-assisted emulsification process. In this work, by mapping the physical processing parameters of ultrasound emulsification into a reduced domain that is governed by the power density and the initial average droplet diameter, a dimensionless parameter that resembles the dynamic breakup potential (η) was established via dimensional analysis. In addition to shedding important insights into the emulsification process, η further facilitates the establishment of a transient scaling relationship that is a function of the characteristic value (a) of the emulsion system. Experimental case study on a cellulose nanocrystals (CNC)-based olein-in-water emulsion system prepared via ultrasound cavitation confirmed the validity of the scaling relationship and sub-universal self-similarity was observed. Using the proposed model, good predictions of the transient of droplet size evolution were attained where the value of η, i.e. the proportionality constant, can be conveniently computed using data from a single time point. Application on other emulsion systems further suggested that the value of a possibly indicates the relative minimum size limit of a particular fluids-emulsifier system. Our approach is general, which encourages widespread adoption for emulsification related studies.
Recently, there have been increasing demand for the application of Pickering emulsions in various industries due to its combined advantage in terms of cost, quality and sustainability. This review aims to provide a complete overview of the available methodology for the physical characterization of emulsions that are stabilized by solid particles (known as Pickering emulsion). Current approaches and techniques for the analysis of the formation and properties of the Pickering emulsion were outlined along with the expected results of these methods on the emulsions. Besides, the application of modelling techniques has also been elaborated for the effective characterization of Pickering emulsions. Additionally, approaches to assess the stability of Pickering emulsions against physical deformation such as coalescence and gravitational separation were reviewed. Potential future developments of these characterization techniques were also briefly discussed. This review can act as a guide to researchers to better understand the standard procedures of Pickering emulsion assessment and the advanced methods available to date to study these emulsions, down to the minute details.
As a class of ionic liquids with higher biocompatibility, cholinium aminoates ([Cho][AA]) hold potential as solvation media for enzymatic bioprocessing. Herein, solvation effect of [Cho][AA] on structural stability and enzymatic activity of Candida antarctica lipase B (CALB) was evaluated using experimental and computational approaches. Influence of [Cho][AA] on CALB stability was investigated using amino acid anions ([AA]-) with varying hydrophobicity levels. Choline phenylalaninate ([Cho][Phe]) resulted in 109.1% and 110.4% of relative CALB activity to buffer medium at 25 °C and 50 °C, respectively. Simulation results revealed the improvement of CALB's enzymatic activities by [AA]- with a strong hydrophobic character. Shielding of CALB from water molecules by [AA]- was observed. The level of CALB activity was governed by accumulation level of [AA]- at CALB's first hydration layer. The stronger interaction between His224 and Asp187 was postulated to be driven by [Cho][AA], resulting in the activity enhancement of CALB. The slight improvement of CALB activity in 0.05 M [Cho][Phe] at 50 °C could be due to the larger size of entrance to the catalytic site and the stronger interaction between the catalytic residues. The promising effect of [Cho][Phe] on CALB activation may stimulate research efforts in designing a 'fully green' bioreaction for various industrial applications.Communicated by Ramaswamy H. Sarma.
Nano-magnetites are widely researched for its potential as an excellent adsorbent in many applications. However, the efficiency of the nano-magnetites are hindered by their tendency to agglomerate. In this work, we dispersed and embedded the nano-magnetites in a porous silica gel matrix to form a nanocomposite to reduce the extent of agglomeration and to enhance the adsorption performance. Our experimental results showed that the removal efficiency of Cu2+ ion has improved by 46% (22.4 ± 2.2%) on the nano-magnetite-silica-gel (NMSG) nanocomposite as compared to pure nano-magnetites (15.3 ± 0.6%). The adsorption capacity is further enhanced by 39% (from 11.2 ± 1.1 to 15.6 ± 1.6 mg/g) by subjecting the NMSG to a magnetic field prior to adsorption. We infer that the magnetic field aligned the magnetic domains within the nano-magnetites, resulting in an increased Lorentz force during adsorption. Similar alignment of magnetic domains is near to impossible in pure nano-magnetites due to severe agglomeration. We further found that the adsorption capacity of the NMSG can be manipulated with an external magnetic field by varying the strength and the configurations of the field. Equipped with proper process design, our finding has great potentials in processes that involve ion-adsorptions, for example, NMSG can: (i) replace/reduce chemical dosing in controlling adsorption kinetics, (ii) replace/reduce complex chemicals required in ion-chromatography columns, and (iii) reduce wastage of nano-adsorbents by immobilizing it in a porous matrix.