Incubation pH Values Research Articles

Saccharification of Various Wastepaper Materials by Cellulase from Brown Garden Snail (Cornu aspersum) at Different Incubation pH Values

Increased solid waste pollution and the negative effect of fossil fuel consumption on the environment are issues that would require more scientific attention and application to deal effectively with these phenomena. Wastepaper, a major component of solid waste, is classified as organic waste due to the presence of cellulose, a glucose-based biopolymer that is part of its structural composition. The saccharification of cellulose into glucose, a fermentable sugar, can be achieved with a hydrolytic enzyme known as cellulase. Although cellulase from fungal species such as Trichoderma, Aspergillus, and Penicillium are well described, knowledge about cellulase isolated from the brown garden snail is limited as it has not been the subject of many research endeavors. The waste paper has been described as a suitable resource for bio-energy development due to cellulose, a structural component of this bio-material that can be degraded into glucose, a fermentable sugar. Although paper materials such as newspaper, office paper, filter paper, Woolworths and Pick and Pay (retailers) advertising paper, as well as foolscap paper, were saccharified by different cellulases, the degradation of these paper materials by garden snail cellulase is a novel investigation from our laboratory. With the effects of temperature and incubation time on this cellulase action when degraded paper materials have already been investigated and reported, this study dealt with the garden snail cellulase action when degraded paper materials at different pH values. Most of the paper materials were degraded optimally at a pH value of 6.0, while optimum saccharification was observed at pH 4.5 when newspaper and brown envelope paper were degraded, with office paper showing maximum bioconversion at pH 7.0. The difference in the structural composition of the paper materials also affects the degree of saccharification, as the amount of sugar released from the various paper materials at optimum pH values is not similar. Together with other catalytic parameters, the pH value of this enzymatic catalysis is also to be considered when designing the development of waste paper as a bio-product resource, with limiting environmental pollution as an additional advantage of this process.

Stable and selective self-assembly of α-lipoic acid on Ge(001) for biomolecule immobilization

We demonstrate a novel method for the stable and selective surface functionalization of germanium (Ge) embedded in silicon dioxide. The Ge(001) surface is functionalized using α-lipoic acid (ALA), which can potentially be utilized for the immobilization of a wide range of biomolecules. We present a detailed pH-dependence study to establish the effect of the incubation pH value on the adsorption layer of the ALA molecules. A threshold pH value for functionalization is identified, dividing the examined pH range into two regions. Below a pH value of 7, the formation of a disordered ALA multilayer is observed, whereas a stable well-ordered ALA mono- to bi-layer on Ge(001) is achieved at higher pH values. Furthermore, we analyze the stability of the ALA layer under ambient conditions, revealing the most stable functionalized Ge(001) surface to effectively resist oxidation for up to one week. Our established functionalization method paves the way towards the successful immobilization of biomolecules in future Ge-based biosensors.

Poly(3,6-diamino-9-ethylcarbazole) based molecularly imprinted polymer sensor for ultra-sensitive and selective detection of 17-β-estradiol in biological fluids

In this work, we reported the synthesis of 3, 6-diamino-9-ethylcarbazole and its application as a new monomer for preparation of molecularly imprinted polymer (MIP) electrochemical sensor. The as prepared MIP sensor exhibited ultrahigh sensitivity and selectivity for the detection of 17-β-estradiol in attomolar levels (1 × 10–18molL−1). The sensor works by detecting the change of the interfacial impedance that is derived from recognition of 17-β-estradiol on the MIP layer. The MIP sensor based on 3, 6-diamino-9-ethylcarbazole monomer revealed better performance than that of unmodified carbazole monomer. The monomer/template ratio, electropolymerization scanning cycles, and the incubation pH values were optimised in order to obtain the best detection efficiency. Under the optimised condition, the MIP sensor exhibits a wide linear range from 1aM to 10μM (1 × 10–18 ̶ 1 × 10−5molL−1). A low detection limit of 0.36aM (3.6 × 10–19molL−1) and a good selectivity towards structurally similar compounds were obtained. The proposed MIP sensor also exhibits long-term stability and applicability in human serum samples. These advantages enabled this MIP sensor to be a promising alternative of electrochemical sensor and may be extended to detection of other endogenous compounds.

Immobilization of adenosine deaminase onto agarose and casein.

In the present study adenosine deaminase (ADA) was immobilized onto two different polymeric materials, agarose and casein. The factors affecting the amount of enzyme attachment onto the polymeric supports such as incubation time were investigated. The maximum amount of enzyme immobilized onto different polymeric supports occurred at incubation pH value 7.5 and ADA concentration 42 units/g and the incubation time needed for the maximum amount of enzyme attachment to the polymeric supports was found to be 8 h. Some phsicochemical properties of the free and immobilized ADA such as operational stability, optimum temperature and thermal stability, pH optimum and stability, storage stability, and the effect of gamma-radiation were studied. The operational stability of the free and immobilized enzyme showed that the enzyme immobilized by a cross-linking technique using gultaric dialdehyde showed poor durability and the relative activity decreased sharply due to the leakage after repeated washing, while the enzymes immobilized by covalent bonds to the carriers showed a slight decrease in most cases in the relative activity (around 20%) after being used 10 times. Storage for 4-6 months, showed that the free enzyme lost its activity, while the immobilized enzyme showed the opposite behavior. Subjecting the immobilized enzyme to a dose of gamma radiation of 0.5-10 Mrad showed complete loss in the activity of the free enzyme at a dose of 5 Mrad, while the immobilized enzymes showed relatively high resistance to gamma radiation up to a dose of 5 Mrad.