Engineering strategies and applications of cyanobacterial exopolysaccharides: A review on past achievements and recent perspectives.

Cyanobacteria are ideally suited for developing sustainable biological products but are underdeveloped due to a lack of genetic tools. Exopolysaccharide (EPS) is one of the essential bioproducts with widespread industrial applications. Despite their unique structural characteristics associated with distinct biological and physicochemical aspects, EPS from cyanobacteria has been underexplored. However, it is expected to accelerate in the near future due to the utilization of low-cost cyanobacterial platforms and readily available information on the structural data and specific features of these biopolymers. In recent years, cyanobacterial EPSs have attracted growing scientific attention due to their simple renewability, rheological characteristics, massive production, and potential uses in several biotechnology domains. This review focuses on the most recent research on potential new EPS producers and their distinct compositions responsible for novel biological activities. Additionally, nutritional and process parameters discovered recently for enhancing EPS production and engineering strategies applied currently to control the biosynthetic pathway for enhanced EPS production are critically highlighted. The process intensification of previously developed EPS extraction and purification processes from cyanobacterial biomass is also extensively explained. Furthermore, the newly reported biotechnological applications of cyanobacterial exopolysaccharides are also discussed.

Astaxanthin from microalgae: A review on structure, biosynthesis, production strategies and application.

Astaxanthin is a red-colored secondary metabolite with excellent antioxidant properties, typically finds application as foods, feed, cosmetics, nutraceuticals, and medications. Astaxanthin is usually produced synthetically using chemicals and costs less as compared to the natural astaxanthin obtained from fish, shrimps, and microorganisms. Over the decades, astaxanthin has been naturally synthesized from Haematococcus pluvialis in commercial scales and remains exceptional, attributed to its higher bioactive properties as compared to synthetic astaxanthin. However, the production cost of algal astaxanthin is still high due to several bottlenecks prevailing in the upstream and downstream processes. To that end, the present study intends to review the recent trends and advancements in astaxanthin production from microalgae. The structure of astaxanthin, sources, production strategies of microalgal astaxanthin, and factors influencing the synthesis of microalgal astaxanthin were discussed while detailing the pathway involved in astaxanthin biosynthesis. The study also discusses the relevant downstream process used in commercial scales and details the applications of astaxanthin in various health related issues.

Comprehensive Review on Biomaterials and Their Inherent Behaviors for Hip Repair Applications.

Developing biomaterials for hip prostheses is challenging and requires dedicated attention from researchers. Hip replacement is an inevitable and remarkable orthopedic therapy for enhancing the quality of patient life for those who have arthritis as well as trauma. Generally, five types of hip replacement procedures are successfully performed in the current medical market: total hip replacements, hip resurfacing, hemiarthroplasty, bipolar, and dual mobility systems. The average life span of artificial hip joints is about 15 years, and several studies have been conducted over the last 60 years to improve the performance and thereby increase the lifespan of artificial hip joints. Present-day prosthetic hip joints are linked to the wide availability of biomaterials. Metals, ceramics, and polymers are some of the most promising types of biomaterials; nevertheless, each biomaterial has advantages and disadvantages. Metals and ceramics fail in most applications owing to stress shielding and the emission of wear debris; ongoing research is being carried out to find a remedy to these unfavorable responses. Recent research found that polymers and composites based on polymers are significant alternative materials for artificial joints. With growing research and several biomaterials, recent reviews lag in effectively addressing hip implant materials' individual mechanical, tribological, and physiological behaviors. This Review comprehensively investigates the historical evolution of artificial hip replacement procedures and related biomaterials' mechanical, tribological, and biological characteristics. In addition, the most recent advances are also discussed to stimulate and guide future researchers as they seek more effective methods and synthesis of innovative biomaterials for hip arthroplasty application.

Nutrient modulation based process engineering strategy for improved butanol production from Clostridium acetobutylicum.

The present study demonstrates a process engineering strategy to achieve high butanol titer and productivity from wild type Clostridium acetobutylicum MTCC 11274. In the first step, two different media were optimized with the objectives of maximizing the biomass and butanol productivity, respectively. In the next step, attributes of these two media compositions were integrated to design a two-stage fed-batch process which resulted in maximal butanol productivity of 0.55 g L-1 h-1 with titer of 13.1 g L-1 . Further, two-stage fed-batch process along with combinatorial use of magnesium limitation and calcium supplementation resulted in the highest butanol titer and productivity of 16.5 g L-1 and 0.59 g L-1 h-1 , respectively. Finally, integration of the process with gas stripping and modulation of feeding duration resulted in a cumulative butanol titer of 54.3 g L-1 and productivity of 0.58 g L-1 h-1 . The strategy opens up possibility of developing a viable butanol bioprocess. © 2019 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2771, 2019.

Real-time iTRAQ-based proteome profiling revealed the central metabolism involved in nitrogen starvation induced lipid accumulation in microalgae.

To understand the post-transcriptional molecular mechanisms attributing to oleaginousness in microalgae challenged with nitrogen starvation (N-starvation), the longitudinal proteome dynamics of Chlorella sp. FC2 IITG was investigated using multipronged quantitative proteomics and multiple reaction monitoring assays. Physiological data suggested a remarkably enhanced lipid accumulation with concomitant reduction in carbon flux towards carbohydrate, protein and chlorophyll biosynthesis. The proteomics-based investigations identified the down-regulation of enzymes involved in chlorophyll biosynthesis (porphobilinogen deaminase) and photosynthetic carbon fixation (sedoheptulose-1,7 bisphosphate and phosphoribulokinase). Profound up-regulation of hydroxyacyl-ACP dehydrogenase and enoyl-ACP reductase ascertained lipid accumulation. The carbon skeletons to be integrated into lipid precursors were regenerated by glycolysis, ß-oxidation and TCA cycle. The enhanced expression of glycolysis and pentose phosphate pathway enzymes indicates heightened energy needs of FC2 cells for the sustenance of N-starvation. FC2 cells strategically reserved nitrogen by incorporating it into the TCA-cycle intermediates to form amino acids; particularly the enzymes involved in the biosynthesis of glutamate, aspartate and arginine were up-regulated. Regulation of arginine, superoxide dismutase, thioredoxin-peroxiredoxin, lipocalin, serine-hydroxymethyltransferase, cysteine synthase, and octanoyltransferase play a critical role in maintaining cellular homeostasis during N-starvation. These findings may provide a rationale for genetic engineering of microalgae, which may enable synchronized biomass and lipid synthesis.

Substrate dependent modulation of butanol to ethanol ratio in non-acetone forming Clostridium sporogenes NCIM 2918.

Present study reports a non-acetone producing Clostridium sporogenes strain as a potential producer of liquid biofuels. Alcohol production was positively regulated by sorbitol and instant dry yeast as carbon and nitrogen sources respectively. Media optimization resulted in maximum butanol and ethanol titer (gL-1) of 12.1 and 7.9 respectively. Depending on the combination of carbon sources, the organism was found to manipulate its metabolism towards synthesis of either ethanol or butanol, thereby affecting the total alcohol titer. Among various dual substrate combinations, glucose-glycerol mixture in the ratio of 60:40 resulted in maximum butanol and ethanol titer (gL-1) of 11.9 and 12.1 respectively with total alcohol productivity of 0.59gL-1h-1. In the mixture, when pure glycerol was replaced with crude glycerol, butanol and ethanol titer (gL-1) of 11.2 and 11.7 was achieved. Hence, the strain shows immense potential for biofuels production using crude glycerol as cheap substrate.

Continuous cultivation of lipid rich microalga Chlorella sp. FC2 IITG for improved biodiesel productivity via control variable optimization and substrate driven pH control.

A novel two-stage continuous heterotrophic cultivation of Chlorella sp. FC2 IITG was demonstrated for enhanced lipid productivity. Initially, effect of control variable e.g. dilution rate and feed stream substrate concentrations on biomass productivity was evaluated. This showed significant variation in biomass productivity from 2.4gL-1day-1 to 11.2gL-1day-1. Further, these control variables were optimized by using multi-nutrient mechanistic model for maximizing the biomass productivity. Finally, continuous production of lipid rich algal biomass was demonstrated in two sequential bioreactors for enhanced lipid productivity. The biomass productivity of 92.7gL-1day-1 was observed in the first reactor which was operated at model predicted optimal substrate concentrations of feed stream. The intracellular neutral lipid enrichment by acetate addition resulted in lipid productivity of 9.76gL-1day-1 in the second reactor. Both the biomass and lipid productivities obtained from current study are significantly high amongst similarly reported literatures.

Synchronized growth and neutral lipid accumulation in Chlorella sorokiniana FC6 IITG under continuous mode of operation.

Synchronized growth and neutral lipid accumulation with high lipid productivity under mixotrophic growth of the strain Chlorella sorokiniana FC6 IITG was achieved via manipulation of substrates feeding mode and supplementation of lipid elicitors in the growth medium. Screening and optimization of lipid elicitors resulted in lipid productivity of 110.59mgL(-1)day(-1) under the combined effect of lipid inducers sodium acetate and sodium chloride. Fed-batch cultivation of the strain in bioreactor with intermittent feeding of limiting nutrients and lipid inducer resulted in maximum biomass and lipid productivity of 2.08 and 0.97gL(-1)day(-1) respectively. Further, continuous production of biomass with concomitant lipid accumulation was demonstrated via continuous feeding of BG11 media supplemented with lipid inducers sodium acetate and sodium chloride. The improved biomass and lipid productivity in chemostat was found to be 2.81 and 1.27gL(-1)day(-1) respectively operated at a dilution rate of 0.54day(-1).

Preferential utilization of intracellular nutrients supports microalgal growth under nutrient starvation: multi-nutrient mechanistic model and experimental validation.

Microalgae are able to grow even under exhaustion of some key nutrients such as nitrogen and phosphorous. Here, we report a multi-nutrient mechanistic model to predict heterotrophic growth of Chlorella sp. FC2 IITG over two sequential phases of fermentation: nutrient sufficient condition to nutrient starved condition. The model assumes that the growth of the microorganism takes place via sequential utilization of extracellular nutrients (ECN) under nutrient replete condition followed by intracellular stored nutrients under exhaustion of limiting nutrients. Further, intracellular nutrient was assumed to be in three different forms: structural form of nutrient (SFN), readily utilizable nutrient (RUN) and non-readily utilizable nutrient (Non-RUN). After the exhaustion of ECN, microorganism switches to RUN followed by Non-RUN to continue its growth, which was experimentally validated by extracting intracellular nitrate and phosphate compounds. The model also incorporates variability in yield coefficients for nitrate and phosphate utilizations.

High cell density lipid rich cultivation of a novel microalgal isolate Chlorella sorokiniana FC6 IITG in a single-stage fed-batch mode under mixotrophic condition.

A single-stage mixotrophic cultivation strategy was developed with a novel microalgal isolate Chlorella sorokiniana FC6 IITG for high cell density lipid-rich biomass generation. The strain was evaluated for growth and lipid content under different physico-chemical parameters, nutritional conditions and trophic modes. Finally, a single-stage mixotrophic fed-batch cultivation strategy was demonstrated with intermittent feeding of key nutrients along with dynamic increase in light intensity for high cell density biomass and sodium acetate as elicitor for lipid enrichment. The key findings: (i) glucose and sodium acetate was identified as growth supporting and lipid inducing nutrients, respectively; (ii) mixotrophic batch cultivation resulted in maximum biomass and lipid productivity (mgL(-1)day(-1)) of 455.5 and 111.85, respectively; (iii) single-stage mixotrophic fed-batch cultivation showed maximum biomass productivity of 1.93gL(-1)day(-1) (biomass titer 15.81gL(-1)) and lipid productivity of 550mgL(-1)day(-1); (iv) biodiesel properties were in accordance with international standards.

Evaluation of indigenous microalgal isolate Chlorella sp. FC2 IITG as a cell factory for biodiesel production and scale up in outdoor conditions.

The present study reports evaluation of an indigenous microalgal isolate Chlorella sp. FC2 IITG as a potential candidate for biodiesel production. Characterization of the strain was performed under photoautotrophic, heterotrophic, and mixotrophic cultivation conditions. Further, an open-pond cultivation of the strain under outdoor conditions was demonstrated to evaluate growth performance and lipid productivity under fluctuating environmental parameters and in the presence of potential contaminants. The key findings were: (1) the difference in cultivation conditions resulted in significant variation in the biomass productivity (73-114 mg l⁻¹ day⁻¹) and total lipid productivity (35.02-50.42 mg l⁻¹ day⁻¹) of the strain; (2) nitrate and phosphate starvation were found to be the triggers for lipid accumulation in the cell mass; (3) open-pond cultivation of the strain under outdoor conditions resulted in biomass productivity of 44 mg l⁻¹ day⁻¹ and total lipid productivity of 10.7 mg l⁻¹ day⁻¹; (4) a maximum detectable bacterial contamination of 7 % of the total number of cells was recorded in an open-pond system; and (5) fatty acid profiling revealed abundance of palmitic acid (C16:0), oleic acid (C18:1) and linoleic acid (C18:2), which are considered to be the key elements for suitable quality biodiesel.

Flux balance analysis of Chlorella sp. FC2 IITG under photoautotrophic and heterotrophic growth conditions.

Quantification of carbon flux distribution in the metabolic network of microalgae remains important to understand the complex interplay between energy metabolism, carbon fixation, and assimilation pathways. This is even more relevant with respect to cyclic metabolism of microalgae under light-dark cycle. In the present study, flux balance analysis (FBA) was carried out for an indigenous isolate Chlorella sp. FC2 IITG under photoautotrophic and heterotrophic growth conditions. A shift in intracellular flux distribution was predicted during transition from nutrient sufficient phase to nutrient starvation phase of growth. Further, dynamic flux analysis (dFBA) was carried out to capture light-dark metabolism over discretized pseudo steady state time intervals. Our key findings include the following: (i) unlike heterotrophic condition, oxidative pentose phosphate (PP) pathway, and Krebs cycle were relatively inactive under photoautotrophic growth; (ii) in both growth conditions, while transhydrogenation reaction was highly active, glyoxalate shunt was found to be nonoperative; (iii) flux distribution during transition period was marked with up regulation of carbon flux toward nongrowth associated (NGA) maintenance energy, oxidative phosphorylation, and photophosphorylation; (iv) redirection of carbon flux from polysaccharide and neutral lipid resulted in up regulation of Krebs cycle flux in the dark phase; (v) elevated glycolytic and acetyl-CoA flux were coupled with induction of neutral lipid during light cycle of the growth; (vi) significantly active photophosphorylation in the light phase was able to satisfy cellular energy requirement without need of oxidative PP pathway; and (vi) unlike static FBA, dFBA predicted an unaltered NGA maintenance energy of 1.5 mmol g(-1) DCW h(-1).