Production of Alginate Oligosaccharides (AOSs) Using Enhanced Physicochemical Properties of Immobilized Alginate Lyase for Industrial Application.

Alginate lyase (AL) is a polysaccharide-degrading enzyme that can degrade alginate by hydrolyzing glycosidic bonds and produces unsaturated alginate oligosaccharides (AOSs). These AOSs have wide therapeutic and nutraceutical applications. However, to produce alginate oligosaccharides in a cost-effective manner is challenging due to the low availability and high cost of this degrading enzyme. Immobilization of the enzyme facilitates industrial applications owing to its stability, reusability, and cost-effectiveness. This study was focused on the enhancement of the properties of alginate lyase and improvement of the production of AOS. Alginate lyase was immobilized on magnetic nanoparticles (NPs) using glutaraldehyde as the crosslinker. The study showed that the maximum binding achieved between NPs and protein in the enzyme was 71% at a ratio of 1:150 NP:protein. As a result of immobilization, the optimum activity of free enzyme which was obtained at 37 °C and pH 7.4 changed to 45 °C and pH 9. Furthermore, the enzyme was thermostable at 45 °C for 3 h with up to 50% reusability for six consecutive cycles. Storage stability after 15 days showed ~67% relative hydrolysis of alginate. The free alginate lyase (25 IU) showed 76% raw biomass (seaweed) hydrolysis which is higher compared to 63% provided by the immobilized enzyme. As a result of efficient hydrolysis, AOSs with molecular weight profile of 370-1040 kDa were produced and detected using HPLC.

Marine bioactives: from energy to nutrition.

Microalgae have been evaluated as promising resource for biodiesel production, but algal biofuel production is not yet commercially viable, which reflects the high energy costs linked with cultivation, harvesting, and dewatering of algae. As crude oil processing declines, microalgae biorefineries are being considered for producing bioactives such as enzymes, proteins, omega-3 oils, pigments, recombinant products, and vitamins, to offset the costs of biofuel production. We believe that producing algal bioactives through advanced manufacturing pathways, encompassing a biorefinery approach, would be effective, profitable, and economical.

Advances on marine-derived natural radioprotection compounds: historic development and future perspective.

Natural extracts and compounds from marine resources have gained intensive scientific and industry attention for radioprotective activities in the past ten years. However, the marine-derived radioprotectants have been studied against UV-rays, gamma (γ)-rays and X-rays for more than 30 years. This review aims to identify key marine-derived extracts/compounds and their modes of action studied for radioprotective activities from 1986 to 2019. A comprehensive survey was conducted to establish the trend in terms of the publications each year and the countries of origin. A total of 40 extracts and 34 natural compounds showing radioprotective activities against UV-rays, gamma (γ)-rays and X-rays were identified from a range of marine plants and animals. These extracts and compounds are broadly categorized into polysaccharides, phlorotannins, carotenoids and mycosporine-like amino acids (MAAs). Macroalgae and microalgae were found to be the dominant sources of polysaccharides, phlorotannins and carotenoids. MAAs were mainly identified in algae, sponges, sea cucumber and corals that showed significant UV-absorbing activities. A number of radioprotective mechanisms were shown by these compounds, predominantly free radicals scavenging, inhibition of apoptosis, UV-ray absorption and DNA damage-repair signaling pathways. While these bio-discoveries warrant further investigation and development of radioprotective therapeutics, however, the lack of clinical studies is a major obstacle to be tackled in the future. Supplementary Information: The online version contains supplementary material available at 10.1007/s42995-021-00095-x.

Release of encapsulated bioactives influenced by alginate viscosity under in-vitro gastrointestinal model.

The physicochemical properties of alginate can affect the release profile of encapsulated bioactives, but this is poorly understood. The influence of alginate viscosity (low- A1, medium- A2 and high- A3) and molecular weight (kDa) on the release of encapsulated bioactives (seaweed and spirulina powder) was investigated in an in-vitro gastrointestinal (GSI) model. Beads encapsulated with A2 at 1% (w/v) have overall higher release of bioactives (protein, phlorotannins and antioxidants) but A3 at 0.5% (w/v) was able to release and absorb similar amount of bioactives with ~10% difference with A2. The relative release of protein, phlorotannins and antioxidant was 96%, 111% and 43% respectively from A2 in gastric digestion. In contrast, protein (165%) and phlorotannins (234%) release was highest from A3 in intestinal phase. These results establish the importance of physicochemical properties of the encapsulating matrix on water retention capacity and their interaction with bioactive material to release into the system.

Seaweed and seaweed-derived metabolites as prebiotics.

Seaweeds and their bioactive compounds, particularly polysaccharides and phenolics can be regarded as great dietary supplements with gut health benefits and prebiotics. These components are resistant to digestion by enzymes present in the human gastrointestinal tract, also selectively stimulate the growth of beneficial gut bacteria and the production of fermentation products such as short chain fatty acids. Commonly, the health benefits of seaweed components are assessed by including them in an in vitro anaerobic fermentation system containing human fecal inocula that mimics the environment of the human large bowel. Regarding to the complex interactions between dietary components, gastrointestinal physiological processes, and gut microbiota are difficult to model in vitro. Consequently it is important to follow up the promising in vitro results with in vivo animal or human testing. The aim of this chapter is to have a comprehensive review on the application of seaweeds and seaweed-derived metabolites as prebiotics, and understand the trends, gaps and future directions of both scientific and industrial developments. This work contributes to develop and expand new platform of seaweed utilization for higher-value products, particularly to functional food and nutraceutical industries in order to serve the social demand for health awareness and support economic development.

Nano-immobilized cellulases for biomass processing with application in biofuel production.

Several researches have focused on the enzymatic pretreatment of lignocellulose biomass to produce fermentable sugars that can lead to ethanol production thus facilitating pathways for sustainable biofuel production. Enzymes are fundamental to the pretreatment process, however, are required in larger quantities during pretreatment process thus influencing biofuel production cost. Immobilization of enzymes to a suitable support/matrix could enhance its stability, and reusability thus containing cost. This chapter focuses on developing an advanced technology for immobilizing enzymes to nanomaterials; variety of nanomaterials used for immobilization, nature of enzyme/protein nanomaterial interactions, methods of enzyme immobilization, and factors affecting mode of interaction for achieving hydrolysis of microcrystalline cellulose and natural cellulosic substrate. The binding of enzyme (94%) to a nanomaterial was established by spectroscopy techniques. The kinetics study, conducted at optimum pH (pH 4) and temperature (50°C for free and 60°C immobilized enzyme), exhibited improvement in immobilized enzyme properties. The immobilized enzyme retained up to 50% of its enzyme activity in five consecutive cycles. This chapter advocates the use of nano-immobilized enzymes in biomass hydrolysis for biofuel production.

Enrichment of Cellulosic Waste Hemp (Cannabis sativa) Hurd into Non-Toxic Microfibres.

In this study a largely available lignocellulose feedstock hemp (Cannabis sativa), obtained as an industrial waste, was used for cellulose extraction. The extraction of cellulose microfibres from hemp biomass was conducted by alkaline treatment and an acidification process. The extracted cellulose microfibres were characterised using Fourier-transformed infrared spectroscopy (FTIR), Scanning electron microscopy (SEM), thermogravimetric analysis (TGA) and X-ray diffraction (XRD). The viability of the study was determined by growing human fibroblasts on the preparation which resulted in being non-toxic; indicating its potential in preparing biological scaffolds. Upon enzymatic hydrolysis of the cellulose microfibre using cellulase from Trichoderma reesei, a maximum of 909 mg/g of reducing sugars were obtained, which endorses its suitability for biofuel production.

Omega-3 fatty acid production from enzyme saccharified hemp hydrolysate using a novel marine thraustochytrid strain.

In this work, a newly isolated marine thraustochytrid strain, Schizochytrium sp. DT3, was used for omega-3 fatty acid production by growing on lignocellulose biomass obtained from local hemp hurd (Cannabis sativa) biomass. Prior to enzymatic hydrolysis, hemp was pretreated with sodium hydroxide to open the biomass structure for the production of sugar hydrolysate. The thraustochytrid strain was able to grow on the sugar hydrolysate and accumulated polyunsaturated fatty acids (PUFAs). At the lowest carbon concentration of 2%, the PUFAs productivity was 71% in glucose and 59% in the sugars hydrolysate, as a percentage of total fatty acids. Saturated fatty acids (SFAs) levels were highest at about 49% of TFA using 6% glucose as the carbon source. SFAs of 41% were produced using 2% of SH. This study demonstrates that SH produced from lignocellulose biomass is a potentially useful carbon source for the production of omega-3 fatty acids in thraustochytrids, as demonstrated using the new strain, Schizochytrium sp. DT3.

Suitability of magnetic nanoparticle immobilised cellulases in enhancing enzymatic saccharification of pretreated hemp biomass.

BACKGROUND: Previous research focused on pretreatment of biomass, production of fermentable sugars and their consumption to produce ethanol. The main goal of the work was to economise the production process cost of fermentable sugars. Therefore, the objective of the present work was to investigate enzyme hydrolysis of microcrystalline cellulose and hemp hurds (natural cellulosic substrate) using free and immobilised enzymes. Cellulase from Trichoderma reesei was immobilised on an activated magnetic support by covalent binding and its activity was compared with that of the free enzyme to hydrolyse microcrystalline cellulose and hemp hurds on the basis of thermostability and reusability. RESULTS: Up to 94% protein binding was achieved during immobilisation of cellulase on nanoparticles. Successful binding was confirmed using Fourier transform infrared spectroscopy (FTIR). The free and immobilised enzymes exhibited identical pH optima (pH 4.0) and differing temperature optima at 50°C and 60°C, respectively. The K M values obtained for the free and immobilised enzymes were 0.87 mg/mL and 2.6 mg/mL respectively. The immobilised enzyme retained 50% enzyme activity up to five cycles, with thermostability at 80°C superior to that of the free enzyme. Optimum hydrolysis of carboxymethyl cellulose (CMC) with free and immobilised enzymes was 88% and 81%, respectively. With pretreated hemp hurd biomass (HHB), the free and immobilised enzymes resulted in maximum hydrolysis in 48 h of 89% and 93%, respectively. CONCLUSION: The current work demonstrated the advantages delivered by immobilised enzymes by minimising the consumption of cellulase during substrate hydrolysis and making the production process of fermentable sugars economical and feasible. The activity of cellulase improved as a result of the immobilisation, which provided a better stability at higher temperatures. The immobilised enzyme provided an advantage over the free enzyme through the reusability and longer storage stability properties that were gained as a result of the immobilisation.