Maurycy Daroch

Astaxanthin-Producing Green Microalga Haematococcus pluvialis: From Single Cell to High Value Commercial Products

Many species of microalgae have been used as source of nutrient rich food, feed, and health promoting compounds. Among the commercially important microalgae, Haematococcus pluvialis is the richest source of natural astaxanthin which is considered as “super anti-oxidant.” Natural astaxanthin produced by H. pluvialis has significantly greater antioxidant capacity than the synthetic one. Astaxanthin has important applications in the nutraceuticals, cosmetics, food, and aquaculture industries. It is now evident that, astaxanthin can significantly reduce free radicals and oxidative stress and help human body maintain a healthy state. With extraordinary potency and increase in demand, astaxanthin is one of the high-value microalgal products of the future.This comprehensive review summarizes the most important aspects of the biology, biochemical composition, biosynthesis, and astaxanthin accumulation in the cells of H. pluvialis and its wide range of applications for humans and animals. In this paper, important and recent developments ranging from cultivation, harvest and postharvest bio-processing technologies to metabolic control and genetic engineering are reviewed in detail, focusing on biomass and astaxanthin production from this biotechnologically important microalga. Simultaneously, critical bottlenecks and major challenges in commercial scale production; current and prospective global market of H. pluvialis derived astaxanthin are also presented in a critical manner. A new biorefinery concept for H. pluvialis has been also suggested to guide toward economically sustainable approach for microalgae cultivation and processing. This report could serve as a useful guide to present current status of knowledge in the field and highlight key areas for future development of H. pluvialis astaxanthin technology and its large scale commercial implementation.

BIOCHEMICAL FEATURES AND BIOETHANOL PRODUCTION OF MICROALGAE FROM COASTAL WATERS OF PEARL RIVER DELTA

This study describes identification, cultivation, monitoring of carbohydrate accumulation and bioethanol production from microalgal strains from the coastal waters of Pearl River Delta. Eighteen identified strains belong to the families Chlorellaceae, Scotiellocystoidaceae, Neochloridaceae, Selenastraceae and Scenedesmaceae. Of isolated strains Mychonastes afer PKUAC 9 and Scenedesmus abundans PKUAC 12 were selected for further biomass and ethanol production analysis. Comparison of three cultivation modes (stationary, shaken and aerated) resulted in the highest biomass productivity obtained for aerated cultures that yielded 0.09 g and 0.11 g dry weight per day per litre of medium for M. afer PKUAC 9 and S. abundans PKUAC 12, respectively. Carbohydrate accumulation monitored by FTIR showed that early stationary phase is optimal for biomass harvest. Microalgal biomass was successfully used as a carbohydrate feedstock for fermentative bioethanol production. S. abundans PKUAC 12 was superior feedstock for bioethanol production when pre-treated with the combination of dilute acid treatment and cellulase.

A cold-adapted esterase from psychrotrophic Pseudoalteromas sp. strain 643A

A psychrotrophic bacterium producing a cold-adapted esterase upon growth at low temperatures was isolated from the alimentary tract of Antarctic krill Euphasiasuperba Dana, and classified as Pseudoalteromonas sp. strain 643A. A genomic DNA library of strain 643A was introduced into Escherichiacoli TOP10F’, and screening on tributyrin-containing agar plates led to the isolation of esterase gene. The esterase gene (estA, 621 bp) encoded a protein (EstA) of 207 amino acid residues with molecular mass of 23,036 Da. Analysis of the amino acid sequence of EstA suggests that it is a member of the GDSL-lipolytic enzymes family. The purification and characterization of native EstA esterase were performed. The enzyme displayed 20–50% of maximum activity at 0–20°C. The optimal temperature for EstA was 35°C. EstA was stable between pH 9 and 11.5. The enzyme showed activity for esters of short- to medium-chain (C4 and C10) fatty acids, and exhibited no activity for long-chain fatty acid esters like that of palmitate and stearate. EstA was strongly inhibited by phenylmethylsulfonyl fluoride, 2–mercaptoethanol, dithiothreitol and glutathione. Addition of selected divalent ions e.g. Mg2+, Co2+ and Cu2+ led to the reduction of enzymatic activity and the enzyme was slightly activated (∼30%) by Ca2+ ions.

Purification, characterisation and expression in Saccharomyces cerevisiae of LipG7 an enantioselective, cold-adapted lipase from the Antarctic filamentous fungus Geomyces sp. P7 with unusual thermostability characteristics.

A lipase, LipG7, has been purified from the Antarctic filamentous fungus Geomyces sp. P7 which was found to be cold-adapted and able to retain/regain its activity after heat denaturation. The LipG7 exhibits 100% residual activity following 1h incubation at 100°C whilst simultaneously showing kinetic adaptations to cold temperatures. LipG7 was also found to have industrial potential as an enantioselective biocatalyst as it is able to effectively catalyse the enantioselective transesterification of a secondary alcohol. The LipG7 coding sequence has been identified and cloned using 454 pyrosequencing of the transcriptome and inverse PCR. The LipG7 protein has been heterologously expressed in Saccharomyces cerevisiae BJ5465 and shown to exhibit the same characteristics as the native protein.

Screening, Growth Medium Optimisation and Heterotrophic Cultivation of Microalgae for Biodiesel Production

This article presents a study on screening of microalgal strains from the Peking University Algae Collection and heterotrophic cultivation for biodiesel production of a selected microalgal strain. Among 89 strains, only five were capable of growing under heterotrophic conditions in liquid cultures and Chlorella sp. PKUAC 102 was found the best for the production of heterotrophic algal biodiesel. Composition of the growth medium was optimised using response surface methodology and optimised growth conditions were successfully used for cultivation of the strain in a fermentor. Conversion of algal lipids to fatty acid methyl esters (FAMEs) showed that the lipid profile of the heterotrophically cultivated Chlorella sp. PKUAC 102 contains fatty acids suitable for biodiesel production.

Induction of lipids and resultant FAME profiles of microalgae from coastal waters of Pearl River Delta.

This article presents a study on identification, cultivation and characterisation of microalgal strains from the coastal waters of the Pearl River Delta in Guangdong, China. Thirty-seven identified strains belong to the families: Chlorellaceae, Scotiellocystoidaceae, Scenedesmaceae,Selenastraceae,Micractiniaceae, Coccomyxaceae, Trebouxiaceae and Chlorococcaceae. Of isolated strains, Hindakia PKUAC 169 was selected for lipid induction using two methods: nitrogen starvation and salt stress. After derivatisation of algal lipids through in situ transesterification, lipid profiles of the alga under the two methods were analysed. The results have shown that both lipid yield and fatty acid profiles vary with the methods. Of the two tested methods of inducing lipid production, salt stress yielded three-fold higher lipid productivity than nitrogen starvation. The lipids are predominantly composed of C14-C18 fatty acids, which are favourable for biodiesel production. Moreover, the content of polyunsaturated fatty acids was below the limit of 12% set by EN14214 biodiesel standard.

Microalgae in aquafeeds for a sustainable aquaculture industry

Due to the rapid global expansion of the aquaculture industry, access to key feedstuffs (fishmeal and fish oil) is becoming increasingly limited because of the finite resources available for wild fish harvesting. This has resulted in other sources of feedstuffs being investigated, namely plant origin substitutes for fishmeal and fish oil for aquafeed. Conventional land-based crops have been favored for some applications as substitutes for a portion of the fishmeal, but they can result in changes in the nutritional quality of the fish produced. Microalgae can be regarded as a promising alternative that can replace fishmeal and fish oil and ensure sustainability standards in aquaculture. They have a potential for use in aquaculture as they are sources of protein, lipid, vitamins, minerals, pigments, etc. This comprehensive review summarizes the most important and recent developments of microalgae use as supplement or feed additive to replace fishmeal and fish oil for use in aquaculture. It also reflects the microalgal nutritional quality and digestibility of microalgae-based aquafeed. Simultaneously, safety and regulatory aspects of microalgae feed applications, major challenges on the use microalgae in aquafeed in commercial production, and future research and development perspective are also presented in a critical manner. This review will serve as a useful guide to present current status of knowledge and highlight key areas for future development of a microalgae-based aquafeed industry and overall development of a sustainable aquaculture industry.

Glycosylated yellow laccases of the basidiomycete Stropharia aeruginosa.

Here we describe the identification, purification and characterisation of glycosylated yellow laccase proteins from the basidiomycete fungus Stropharia aeruginosa. Biochemical characterisation of two yellow laccases, Yel1p and Yel3p, show that they are both secreted, monomeric, N-glycosylated proteins of molecular weight around 55kDa with substrate specificities typical of laccases, but lacking the absorption band at 612nm typical of the blue laccase proteins. Low coverage, high throughput 454 transcriptome sequencing in combination with inverse-PCR was used to identify cDNA sequences. One of the cDNA sequences has been assigned to the Yel1p protein on the basis of identity between the translated protein sequence and the peptide data from the purified protein, and the full length gene sequence has been obtained. Biochemical properties, substrate specificities and protein sequence data have been used to discuss the unusual spectroscopic properties of S. aeruginosa proteins in the context of recent theories about the differences between yellow and blue laccases.

Positive effects of duckweed polycultures on starch and protein accumulation.

The effect of duckweed species composition (Lemna aequinoctialis 5505, Landoltia punctata 5506 and Spirodela polyrhiza 5507) in polyculture and monoculture on biomass and starch/protein content were investigated at different levels of temperature, light intensity, nitrogen and phosphorus concentrations. The three growth parameters significantly affect duckweed biomass accumulation. Different combinations of duckweed species greatly varied in starch/protein content. Although all the polycultures showed a median relative growth rate and the majority of the polycultures showed a median and starch/protein content as compared with their respective monocultures, some of the polycultures were found to promote the accumulation of starch/protein at different growth conditions. These findings indicated that proper combination of duckweed species could facilitate desirable biomass accumulation and improve biomass quality. The present study provides useful references for future large-scale duckweed cultivation.

Effects of pH, initial Pb 2+ concentration, and polyculture on lead remediation by three duckweed species

Various geographical duckweed isolates have been developed for phytoremediation of lead. The Pb2+ removal efficiency of Lemna aequinoctialis, Landoltia punctata, and Spirodela polyrhiza was investigated in monoculture and polyculture at different levels of pH and initial Pb2+ concentrations. L. aequinoctialis was not sensitive to the tested pH but significantly affected by initial Pb2+ concentration, whereas synergistic effect of pH and initial Pb2+ concentration on removal efficiency of L. punctata and S. polyrhiza was found. Although the majority of polycultures showed median removal efficiency as compared to respective monocultures, some of the polycultures achieved higher Pb2+ removal efficiencies and can promote population to remove Pb2+. Besides, the three duckweed strains could be potential candidates for Pb2+ remediation as compared to previous reports. Conclusively, this study provides useful references for future large-scale duckweed phytoremediation.