A Saponification Method for Chlorophyll Removal from Microalgae Biomass as Oil Feedstock.

Microalgae oil is an optimal feedstock for nutraceutical, pharmaceutical and biodiesel production, but its high levels of chlorophyll limit its large-scale application. To date, few effective approaches have been developed to remove chlorophyll from microalgae oil. The main purpose of this study was to present a preprocessing method of algae oil feedstock (Scenedesmus) to remove chlorophyll by saponification. The results showed that 96% of chlorophyll in biomass was removed. High quality orange transparent oil could be extracted from the chlorophyll reduced biomass. Specifically, the proportion of neutral lipids and saturation levels of fatty acids increased, and the pigments composition became carotenoids-based. The critical parameters of chlorophyll reduced biodiesel conformed to the standards of the USA, China and EU. Sodium copper chlorophyllin could be prepared from the bleaching effluent. The results presented herein offer a useful pathway to improve the quality of microalgae oil and reduce the cost of microalgae biodiesel.

Gelatiniphilus marinus gen. nov., sp. nov., a bacterium from the culture broth of a microalga, Picochlorum sp. 122, and emended description of the genus Hwangdonia.

A Gram-stain-negative, non-motile, non-spore-forming, rod-shaped bacterium, designated strain GYP-24T, was isolated from the culture broth of a marine microalga, Picochlorum sp. 122. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain GYP-24T forms a robust cluster with H.wangdoniaseohaensis KCTC 32177T (95.8 % sequence similarity) in the family Flavobacteriaceae. Growth of strain GYP-24T was observed at 15, 22, 28, 30, 33 and 37 °C (optimal 30-33 °C), pH 6.0-10.0 (optimal pH 7.0-8.0) and in the presence of 0.5-4 % (w/v) NaCl (optimal 2-3 %). The only menaquinone of strain GYP-24T was MK-6, and the G+C content of the genomic DNA was 36.9 mol%. The major fatty acid profile comprised iso-C17 : 0 3-OH, summed feature 3 (C16 : 1 ω7c/ω6c), iso-C15 : 1 G and iso-C15 : 0. The major polar lipids of strain GYP-24T were phosphatidylethanolamine, one unidentified phospholipid, three unidentified aminolipids and three unidentified lipids. Comprehensive analyses based on polyphasic characterization of GYP-24T indicated that it represents a novel species of a new genus, for which the name Gelatiniphilus marinus gen. nov., sp. nov. is proposed. The type strain is GYP-24T (=KCTC 42903T=MCCC 1K01730T). An emended description of the genus Hwangdonia is also given.

Pyruvatibacter mobilis gen. nov., sp. nov., a marine bacterium from the culture broth of Picochlorum sp. 122.

A Gram-stain-negative, aerobic bacterium, designated strain GYP-11T, was isolated from the culture broth of a marine microalga, Picochloruma sp. 122. Cells were dimorphic rods; free living cells were motile by means of a single polar flagellum, and star-shaped-aggregate-forming cells were attached with stalks and non-motile. Sodium pyruvate or Tween 20 was required for growth on marine agar 2216.16S rRNA gene sequence analysis revealed that this isolate shared 94.07 % similarity with its closest type strain, Parvibaculum hydrocarboniclasticum EPR92T. Phylogenetic analyses indicated that strain GYP-11T represents a distinct lineage in a robust clade consisting of strain GYP-11T, alphaproteobacterium GMD21A06 and Candidatus Phaeomarinobacter ectocarpi Ec32. This clade was close to the genera Parvibaculum and Tepidicaulis in the order Rhizobiales. Chemotaxonomic and physiological characteristics, including cellular fatty acids and carbon source profiles, also readily distinguished strain GYP-11T from all established genera and species. Thus, it is concluded that strain GYP-11T represents a novel species of a new genus in the order Rhizobiales, for which the name Pyruvatibacter mobilis gen. nov., sp. nov. is proposed. The type strain of Pyruvatibacter mobilis is GYP-11T ( = CGMCC 1.15125T = KCTC 42509T).

Aliikangiella marina gen. nov., sp. nov., a marine bacterium from the culture broth of Picochlorum sp. 122, and proposal of Kangiellaceae fam. nov. in the order Oceanospirillales.

A Gram-stain-negative, non-motile, non-spore-forming, long rod-shaped bacterium, designated strain GYP-15T, was isolated from the culture broth of a marine microalga, Picochloruma sp. 122. Phylogenetic analyses revealed that strain GYP-15T shared 90.6 % 16S rRNA gene sequence similarity with its closest relative, Kangiella aquimarina KCTC 12183T, and represents a distinct phylogenetic lineage in a robust clade consisting of GYP-15T and members of the genera Kangiella and Pleionea in the order Oceanospirillales. Chemotaxonomic and physiological characteristics, including major cellular fatty acids, NaCl tolerance and pattern of carbon source utilization, could also readily distinguish strain GYP-15T from all established genera and species. Thus, it is concluded that strain GYP-15T represents a novel species of a new genus, for which the name Aliikangiella marina gen. nov., sp. nov. is proposed. The type strain of Aliikangiella marina is GYP-15T ( = MCCC 1K01163T = KCTC 42667T). Based on phylogenetic results, 16S rRNA gene signature nucleotide pattern and some physiological characteristics, the three genera Kangiella, Pleionea and Aliikangiella are proposed to make up a novel family, Kangiellaceae fam. nov., in the order Oceanospirillales.

Maribacter flavus sp. nov., isolated from a cyanobacterial culture pond.

A Gram-stain-negative, yellow, non-spore-forming, strictly aerobic bacterium, designated C3T, was isolated from a cyanobacterial culture pond. Cells were halophilic, rod-shaped and able to move by gliding. Growth of strain C3T was observed at 15-30 °C (optimum 25 °C), pH 6.0-9.0 (optimum pH 7.5), and in the presence of 1-7 % (w/v) NaCl (optimum 2-3 %). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain C3T formed a distinct lineage within the family Flavobacteriaceae and exhibited the highest similarity (95.21 %) to the type strains of Maribacter dokdonensis, Maribacter arcticus, Maribacter orientalis and Maribacter stanieri, and 'Maribacter caenipelagi' HD-44. The only isoprenoid quinone present within strain C3T was menaquinone 6 (MK-6). The G+C content of genomic DNA was 41.5 mol%. The major polar lipids were phosphatidylethanolamine and three unidentified lipids. The predominant cellular fatty acids (>5 % of the total fatty acids) were iso-C15 : 1 G, iso-C15 : 0, iso-C15 : 0 3-OH, iso-C17 : 0 3-OH, and summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c). On the basis of the phenotypic, chemotaxonomic and phylogenetic characteristics, strain C3T represents a novel species of the genus Maribacter, for which the name Maribacter flavus sp. nov. is proposed. The type strain is C3T ( = KCTC 42508T = CGMCC 1.15112T).

The influence of cultivation period on growth and biodiesel properties of microalga Nannochloropsis gaditana 1049.

This work reported for the first time the detailed impacts of cultivation period on growth dynamics and biochemical composition of a microalga strain Nannochloropsis gaditana 1049. The results shown either the biomass accumulation, lipid content, neutral lipid content, monounsaturated fatty acids composition or the favorable fatty acid profile of C16-C18 increased along with the cultivation period extension, but the lipid productivity displayed a decrease since cultured for 16 days, with the highest value reached 289.51 ± 16.34 mg L(-1) d(-1). Biodiesel properties of this microalga also changed with the cultivation period extension, with average unsaturated degree decreased from 1.24 ± 0.03 to 0.59 ± 0.02, cloud point increased from 3.39 ± 0.40 °C to 12.14 ± 0.32 °C, cetane number increased from 54.59 ± 0.20 to 58.96 ± 0.16 and iodine number reduced sharply from 105.15 ± 2.24 gI2/100g to 56.44 ± 1.76 gI2/100g, which all satisfied the specifications of biodiesel standard.

Wenzhouxiangella marina gen. nov, sp. nov, a marine bacterium from the culture broth of Picochlorum sp. 122, and proposal of Wenzhouxiangellaceae fam. nov. in the order Chromatiales.

A Gram-stain negative, non-motile, non-phototrophic, non-alkaliphilic, obligately aerobic, chemoheterotrophic, and rod-shaped bacterium, designated strain Ma-11(T), was isolated from the culture broth of a marine microalga, Picochloruma sp. 122. Phylogenetic analyses showed that strain Ma-11(T) has less than 91 % similarity to its closest relative, Thioalkalivibrio sulfidiphilus HL-EbGR7(T), represents a distinct phylogenetic lineage in the order Chromatiales, and could not be assigned to any defined families in this order. Chemotaxonomic, genetic and physiological characteristics, including major fatty acids, genomic G+C content, lack of motility, aerophilicity and chemoheterotrophicity, could readily distinguish strain Ma-11(T) from any established members of the order Chromatiales. Based on the 16S rRNA gene sequence analysis and its signature nucleotide pattern, a new family Wenzhouxiangellaceae fam. nov. comprising the genus Wenzhouxiangella gen. nov. and species Wenzhouxiangella marina sp. nov. is proposed. The type strain is Ma-11(T) (=CGMCC 1.14936(T) = KCTC 42284(T) = MCCC 1K00261(T)).

Bacterioplanes sanyensis gen. nov., sp. nov., a PHB-accumulating bacterium isolated from a pool of Spirulina platensis cultivation.

A Gram-negative, poly-3-hydroxybutyrate-accumulating rod bacterium, strain GYP-2(T), was isolated from a pool of marine Spirulina platensis cultivation, Sanya, China. Growth was observed at 10-45 °C and pH 6-10 in the presence of 1-10 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the new isolate belonged to Gammaproteobacteria and displayed 93.8-95.3 % 16S rRNA gene sequences similarities to members of the genera Thalassolituus, Oleibacter, and Oceanobacter, but house-keeping gene gyrB (encode DNA gyrase beta subunit) demonstrated that the new isolate was distantly related to Thalassolituus, Oleibacter, and Oceanobacter species (only 77-83 % gene gyrB sequences similarities).The G+C content of genomic DNA was 55 mol%. The major respiratory quinone was Q-9, while that for Oceanobacter kriegii LMG 6238(T) was Q-8. Major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylethanolamine. On the basis of its physiological, chemotaxonomic, and molecular properties, strain GYP-2(T) is suggested to represent a novel species of a new genus in Gammaproteobacteria, for which the name Bacterioplanes sanyensis gen. nov., sp. nov. is proposed. The type strain is GYP-2(T) (=CGMCC 1.12392(T)=KCTC 32220(T)).

Genome sequences of two pseudoalteromonas strains isolated from the South china sea.

Two Pseudoalteromonas strains, SCSIO 04301 and SCSIO 11900, were isolated from the South China Sea, and both strains form biofilms. Here we present the draft genome sequences of these two strains, which will aid the study of marine microbes that are adapted to marine sediments or are associated with eukaryotic hosts.

Pustulibacterium marinum gen. nov., sp. nov., a member of the family Flavobacteriaceae isolated from the Bashi Channel.

A Gram-reaction-negative, non-spore-forming, gliding, non-translucent, colourless or yellow, aerobic and elevated-colony-forming strain, designated E403(T), was isolated from the Bashi Channel and subjected to a polyphasic taxonomic study. Strain E403(T) could grow in the presence of 0.3-8 % (w/v) NaCl, at 16-43 °C and at pH 6-9, and grew optimally at 28 °C, pH 8, in natural seawater medium. The respiratory quinones were MK-6 and MK-7. The major fatty acids were iso-C15 : 0, iso-C17 : 0 3-OH, iso-C15 : 1 G, summed feature 3 (C16 : 1ω7c/C16 : 1ω6c), iso-C15 : 0 3-OH and C16 : 0. The DNA G+C content of strain E403(T) was 37.9 mol%. Phylogenetic analyses based on 16S rRNA gene sequences of members of the family Flavobacteriaceae showed that strain E403(T) formed a distinct evolutionary lineage within the stable cluster containing type strains Zhouia amylolytica HN-171(T) (92.2 % similarity) and Joostella marina En5(T) (92.4 % similarity). In addition to the large 16S rRNA gene sequence differences, E403(T) can also be distinguished from the reference type strains J. marina En5(T) and Sinomicrobium oceani SCSIO 03483(T) by several phenotypic characteristics and chemotaxonomic properties. On the basis of phenotypic, chemotaxonomic and phylogenetic properties, strain E403(T) is suggested to represent a novel species of a new genus in the family Flavobacteriaceae, for which the name Pustulibacterium marinum gen. nov., sp. nov. is proposed. The type strain is E403(T) (= CCTCC AB2012862(T) = CGMCC 1.12333(T) = KCTC 32192(T)).