Structures of the Apo and FAD-Bound Forms of 2-Hydroxybiphenyl 3-monooxygenase (HbpA) Locate Activity Hotspots Identified by Using Directed Evolution

The FAD-dependent monooxygenase HbpA from Pseudomonas azelaica HBP1 catalyses the hydroxylation of 2-hydroxybiphenyl (2HBP) to 2,3-dihydroxybiphenyl (23DHBP). HbpA has been used extensively as a model for studying flavoprotein hydroxylases under process conditions, and has also been subjected to directed-evolution experiments that altered its catalytic properties. The structure of HbpA has been determined in its apo and FAD-complex forms to resolutions of 2.76 and 2.03 Å, respectively. Comparisons of the HbpA structure with those of homologues, in conjunction with a model of the reaction product in the active site, reveal His48 as the most likely acid/base residue to be involved in the hydroxylation mechanism. Mutation of His48 to Ala resulted in an inactive enzyme. The structures of HbpA also provide evidence that mutants achieved by directed evolution that altered activity are comparatively remote from the substrate-binding site

A reductive aminase from Aspergillus oryzae

Reductive amination is one of the most important methods for the synthesis of chiral amines. Here we report the discovery of an NADP(H)-dependent reductive aminase from Aspergillus oryzae (AspRedAm, Uniprot code Q2TW47) which can catalyse the reductive coupling of a broad set of carbonyl compounds with a variety of primary and secondary amines with up to >98% conversion and with up to >98% enantiomeric excess. In cases where both carbonyl and amine show high reactivity, it is possible to employ a 1:1 ratio of the substrates, forming amine products with up to 94% conversion. Steady-state kinetic studies establish that the enzyme is capable of catalysing imine formation as well as reduction. Crystal structures of AspRedAm in complex with NADP(H) and also with both NADP(H) and the pharmaceutical ingredient (R)-rasagiline are reported. We also demonstrate preparative scale reductive aminations with wild-type and Q240A variant biocatalysts displaying total turnover numbers of up to 32,000 and space time yields up to 3.73 g L-1 d-1

Stereoselectivity and Structural Characterisation of an Imine Reductase (IRED) from Amycolatopsis orientalis

The imine reductase AoIRED from Amycolatopsis orientalis (Uniprot R4SNK4) catalyzes the NADPH-dependent reduction of a wide range of prochiral imines and iminium ions, predominantly with (S)-selectivity and with e.e.s of up to >99%. AoIRED displays up to 100-fold greater catalytic efficiency for 2-methyl-1-pyrroline (2MPN) compared to other IREDs, such as the enzyme from Streptomyces sp. GF3546, which also exhibits (S)-selectivity, and thus AoIRED is an interesting candidate for preparative synthesis. AoIRED exhibits unusual catalytic properties, with inversion of stereoselectivity observed between structurally similar substrates, and also, in the case of 1-methyl-3,4-dihydroisoquinoline, for the same substrate, dependent on the age of the enzyme after purification. The structure of AoIRED has been determined in an ‘open’ apo-form, revealing a canonical dimeric IRED fold in which the active site is formed between the N- and C-terminal domains of participating monomers. Co-crystallisation with NADPH gave a ‘closed’ form in complex with the cofactor, in which a relative closure of domains, and associated loop movements, has resulted in a much smaller active site. A ternary complex was also obtained by co-crystallization with NADPH and 1-methyl-1,2,3,4-tetrahydroisoquinoline [(MTQ], and reveals a binding site for the (R)-amine product which places the chiral carbon within 4 Å of the putative location of the C4 atom of NADPH that delivers hydride to the C=N bond of the substrate. The ternary complex has permitted structure-informed mutation of the active site, resulting in mutants including Y179A, Y179F and N241A, of altered activity and stereoselectivity

Aboriginal Food Security in Northern Canada: An Assessment of the State of Knowledge

As the world’s population increases, as global markets become more interconnected, and as the effects of climate change become clearer, the issue of food insecurity is gaining traction at local, national, and international levels. The recent global economic crisis and increased food prices have drawn attention to the urgent situation of the world’s 870 million chronically undernourished people who face the number one worldwide risk to health: hunger and malnutrition. Although about 75% of the world’s undernourished people live in low-income, rural regions of developing countries, hunger is also an issue in Canada. In 2011, 1.6 million Canadian households, or slightly more than 12%, experienced some level of food insecurity. About one in eight households are affected, including 3.9 million individuals. Of these, 1.1 million are children. Food insecurity presents a particularly serious and growing challenge in Canada’s northern and remote Aboriginal communities (see Figure 1). Evidence from a variety of sources concludes that food insecurity among northern Aboriginal peoples is a problem that requires urgent attention to address and mitigate the serious impacts it has on health and well-being. Results from the 2007–2008 International Polar Year Inuit Health Survey indicate that Nunavut has the highest documented rate of food insecurity for any Indigenous population living in a developed country. According to estimates from the 2011 Canadian Community Health Survey (CCHS), off-reserve Aboriginal households across Canada experience food insecurity at a rate that is more than double that of all Canadian households (27%). Recent data indicate that Canadian households with children have a higher prevalence of food insecurity than households without children, and preliminary evidence indicates that more women than men are affected

Emerging technologies for conversion of sustainable macroalgal carrageenan biomass into L-lactic acid: A state-of-the-art review

The environmental awareness and concerns (plastic pollution) worldwide have driven the development of sustainable and environmentally friendly biopolymer derived from renewable materials. Biopolymers, especially L-lactic acid (L-LA) have played a crucial role in manufacturing polylactic acid, a biodegradable thermoplastic. Recently, L-LA production from non-edible macroalgal biomass has gained immense attraction due to it offers the simplest saccharification process for the biorefinery route. However, the commercialization of macroalgal-based L-LA is still limited due to high production costs. This paper has comprehensively reviewed the potential and development of third-generation feedstock for L-LA production, including significant technological barriers to be overcome for potential commercialization purposes. Then, an insight into the state-of-the-art hydrolysis and fermentation technologies using macroalgae as feedstock are also deliberated in detail. Furthermore, this review provides a conceivable picture of macroalgae-based L-LA biorefinery and future research directions that can be served as an important guideline for scientists, policymakers, and industrial players

A mini-investigation on enhanced oil recovery evolution (2007 – 2020)

Energy plays an important role in sustaining humanity. With rising worldwide energy demand and the great dependence of energy generation on fossil fuels, it is inevitable that enhanced oil recovery must be deployed to recover more possible reserves. This report focuses on reviewing publications related to enhanced oil recovery from 2007 to 2020 through the utilization of bibliometric analysis. Of the 5498 documents retrieved from Web of Science, 569 journals, 90 countries, 2025 organizations, and 8684 authors are involved. China, the United States, Iran, Canada, and India published the most documents. The United States has the highest h-index at 61. The analysis of keywords had shown that the hot issues lie around four main domains namely carbon capture, utilization, and sequestration (CCUS), microbial enhanced oil recovery (MEOR), development of unconventional reserves, and chemical enhanced oil recovery. This study provides some useful insights for future research directions. From there, discussions were subsequently placed on chemical EOR