Unexpected growth promotion of Chlorella sacchrarophila triggered by herbicides DCMU.

The ecotoxicological effects of herbicide contamination on the autotrophic growth of microalgae in aquatic environments have been major concerns. However, little is known about the influence of herbicides on the mixotrophic growth of microalgae. This study investigated the ecotoxicological effect of 3-(3,4-dichlorophenyl)-1,1-dimethyl-urea (DCMU) on the mixotrophic growth of Chlorella sacchrarophila FACHB 4. Results showed that C. sacchrarophila in mixotrophy was more resistant to DCMU than in photoautotrophy. Moreover, a low content of DCMU (20-80 µg·L-1) promoted the mixotrophic growth of C. sacchrarophila, and the promotion effect was obviously enhanced with the increase in light intensity. The chlorophyll content and glucose absorption rate of C. sacchrarophila were found to increase after incubation with DCMU for 24 h. Transcriptome analyses revealed that the mechanism of DCMU to promote the mixotrophic growth of C. sacchrarophila was probably through accelerating glucose uptake and utilization, which was accomplished by reducing photodamage and increasing the chlorophyll content of C. sacchrarophila. This study not only revealed an unexpected bloom of mixotrophic microalgae triggered by herbicides, but it also shed new light on an effective and low-cost strategy to improve the microalgae productivity for utilization.

Co-cultivation of microalga and xylanolytic bacterium by a continuous two-step strategy to enhance algal lipid production.

To enhance microalgal lipid production, canonical two-step cultivation strategy that by transferring the microalgal cells grown in nutrient-replete medium to nutrient-depleted medium is widely used. However, the harvesting step during the transfer raises the production cost. To avoid the harvesting step, this study developed a continuous two-step (CTS) cultivation strategy. In the strategy, Chlorella sacchrarophila was grown in bioreactor while a xylanolytic bacterium Cellvibrio pealriver grown in an inner bag that embedded in the bioreactor; after the first-step co-cultivation, the inner bag is removed which then start the second-step cultivation of C. sacchrarophila. Based on the strategy, the lipid production was determined as 825.34-929.79 mg·L-1, which were 1.7-1.9 times higher than that of cultivation in canonical two-step strategy using glucose as feedstock. During the CTS strategy, the co-cultivation using xylan as feedstock promotes the microalgal growth and the removal of inner bag produces nutrient-depleted condition for enhancing microalgal lipid production.

Parasulfuritortus cantonensis gen. nov., sp. nov., a microaerophilic sulfur-oxidizing bacterium isolated from freshwater sediment.

A novel sulfur-oxidizing bacterium, designated strain LSR1T, was enriched and isolated from a freshwater sediment sample collected from the Pearl River in Guangzhou, PR China. The strain was an obligate chemolithoautotroph, using thiosulfate or sulfide as an electron donor and energy source. Growth of strain LSR1T was observed at 15-40 °C, pH 6.0-7.5 and NaCl concentrations of 0-1.5 %. Strain LSR1T was microaerophilic, with growth only at oxygen content less than 10 %. Anaerobic growth was also observed when using nitrate as the sole electron acceptor. The major cellular fatty acids were C16 : 0 and summed feature 3 (comprising C16 : 1 ω7c and/or C16 : 1 ω6c). The DNA G+C content of the draft genome sequence was 67.5 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain LSR1T formed a lineage within the family Thiobacillaceae, showing sequence identities of 92.87, 92.33 and 90.80 % with its closest relative genera Sulfuritortus, Annwoodia and Thiobacillus, respectively. The genome of strain LSR1T contained multiple genes encoding sulfur-oxidizing enzymes that catalyse thiosulfate and sulfide oxidation, and the gene encoding cbb 3-type cytochrome c oxidase and bd-type quinol oxidase, which enables strain LSR1T to perform sulphur oxidation under microaerophilic conditions. On the basis of phenotypic, genotypic and phylogenetic results, strain LSR1T is considered to represent a novel species of a new genus Parasulfuritortus within the family Thiobacillaceae, for which the name Parasulfuritortus cantonensis gen. nov., sp. nov. is proposed. The type strain is LSR1T (=GDMCC 1.1549=JCM 33645).

Mixotrophic cultivation of Chlorella for biomass production by using pH-stat culture medium: Glucose-Acetate-Phosphorus (GAP).

Here the study designed a pH-stat culture medium that named as Glucose-Acetate-Phosphorus (GAP) for the mixotrophic cultivation of Chlorella for biomass production. With no addition of pH buffer, the culture pH during mixotrophic growth was effectively maintained steady between 7.5 and 8.5 by balancing the ammonium, acetate and glucose uptakes. Based on the GAP medium supplying with 2 g·L-1 of total organic carbon, the biomass productions of four Chlorella species were determined as 4.08-4.56 g·L-1. In contrast to the cultivation using medium Tris-Acetate-Phosphorus (TAP), a algal culture medium that usually regarded as specific for mixotrophy, the cultivation in GAP were about 1.79-1.86 times higher in biomass production and 83.9-88.9% lower in production cost. The developed GAP medium is a promising alternative for the mixotrophic cultivation of microalgae to produce biomass and cellular contents.

The Applications of Promoter-gene-Engineered Biosensors.

A promoter is a small region of a DNA sequence that responds to various transcription factors, which initiates a particular gene expression. The promoter-engineered biosensor can activate or repress gene expression through a transcription factor recognizing specific molecules, such as polyamine, sugars, lactams, amino acids, organic acids, or a redox molecule; however, there are few reported applications of promoter-enhanced biosensors. This review paper highlights the strategies of construction of promoter gene-engineered biosensors with human and bacteria genetic promoter arrays with regard to high-throughput screening (HTS) molecular drugs, the study of the membrane protein's localization and nucleocytoplasmic shuttling mechanism of regulating factors, enzyme activity, detection of the toxicity of intermediate chemicals, and probing bacteria density to improve value-added product titer. These biosensors' sensitivity and specificity can be further improved by the proposed approaches of Mn2+ and Mg2+ added random e error-prone PCR that is a technique used to generate randomized genomic libraries and site-directed mutagenesis approach, which is applied for the construction of bacteria's "mutant library". This is expected to establish a flexible HTS platform (biosensor array) to large-scale screen transcription factor-acting drugs, reduce the toxicity of intermediate compounds, and construct a gene-dynamic regulatory system in "push and pull" mode, in order to effectively regulate the valuable medicinal product production. These proposed novel promoter-engineered biosensors aiding in synthetic genetic circuit construction will maximize the efficiency of the bio-synthesis of medicinal compounds, which will greatly promote the development of microbial metabolic engineering and biomedical science.

Potential of using sodium bicarbonate as external carbon source to cultivate microalga in non-sterile condition.

In this study, a saline-alkaline tolerant microalgal strain was isolated and identified as Chlorella sp. LPF. This strain was able to grow at pH values up to 10 and at salinities up to 5%, and tolerated to 80 g L-1 of sodium bicarbonate. The utilization of bicarbonate as carbon source significantly promoted microalgal growth and lipid production. In the non-sterile cultivation supplying with 80 g L-1 of sodium bicarbonate, the microalgal growth had no difference with their growth in the sterile medium; however, the bacterial growth was suppressed and the cell number decreased to low levels after six days cultivation. This study gives an insight into the potential that using high concentration of sodium bicarbonate as external carbon source to cultivate microalga in non-sterile condition, and suggests a possibility of using bicarbonate as growth promoter and antibacterial agent for the microalgal outdoor cultivation.

Comparative Phenotype and Genome Analysis of Cellvibrio sp. PR1, a Xylanolytic and Agarolytic Bacterium from the Pearl River.

Cellvibrio sp. PR1 is a xylanolytic and agarolytic bacterium isolated from the Pearl River. Strain PR1 is closely related to Cellvibrio fibrivorans and C. ostraviensis (identity > 98%). The xylanase and agarase contents of strain PR1 reach up to 15.4 and 25.9 U/mL, respectively. The major cellular fatty acids consisted of C16:0 (36.7%), C18:0 (8.8%), C20:0 (6.8%), C15:0 iso 2-OH or/and C16:1ω7c (17.4%), and C18:1ω7c or/and C18:1ω6c (6.7%). A total of 251 CAZyme modules (63 CBMs, 20 CEs, 128 GHs, 38 GTs, and 2 PLs) were identified from 3,730 predicted proteins. Genomic analysis suggested that strain PR1 has a complete xylan-hydrolyzing (5 ß-xylanases, 16 ß-xylosidases, 17 α-arabinofuranosidases, 9 acetyl xylan esterases, 4 α-glucuronidases, and 2 ferulic acid esterases) and agar-hydrolyzing enzyme system (2 ß-agarases and 2 α-neoagarooligosaccharide hydrolases). In addition, the main metabolic pathways of xylose, arabinose, and galactose are established in the genome-wide analysis. This study shows that strain PR1 contains a large number of glycoside hydrolases.

Promotion of microalgal growth by co-culturing with Cellvibrio pealriver using xylan as feedstock.

In this work, a Cellvibrio pealriver-microalga co-cultivation mode was used to promote the growths of four microalgae by using xylan as feedstock. After 12days of cultivation, the biomass concentrations of Chlorella sacchrarophila, Chlorella pyrenoidosa and Chlamydomonas reinhardtii in co-cultivation were equal to those in mixotrophic growth on glucose, and the Dunaliella was about 1.6-fold higher than that on glucose. The comparative transcriptomes analysis demonstrated that the xylose and xylan hydrolysates were catalyzed to some active substrates by C. pealriver via some functional enzymes; these active substrates are possibly responsible for the promotion of microalgal growth. This C. pealriver-microalga co-cultivation mode is a potential method to produce low-cost microalgal biodiesel by using hemicellulose as feedstock.

Genome sequence of Cellvibrio pealriver PR1, a xylanolytic and agarolytic bacterium isolated from freshwater.

Cellvibrio pealriver PR1 (CGMCC 1.14955=NBRC 110968) was isolated from a freshwater sample from the Pearl River in China. It is able to degrade various carbohydrates such as starch, xylan, agar, cellulose or chitin. The genomic feature and polysaccharide hydrolases of this strain were described in this paper. The total genome size of C. pealriver PR1 is 4,427,922 bp with 3986 coding sequences (CDS), 53 tRNAs, 16 rRNAs and 1 sRNA. The annotated full genome sequence of this strain provides the genetic basis for revealing its role as a xylanolytic and agarolytic bacterium.