Astaxanthin preparation by fermentation of esters from Haematococcus pluvialis algal extracts with Stenotrophomonas species

Natural astaxanthin (Ax) is an additive that is widely used because of its beneficial biochemical functions. However, the methods used to produce free Ax have drawbacks. Chemical saponification methods produce several by‐products, and lipase‐catalyzed hydrolysis methods are not cost effective. In this study, a bacterial strain of Stenotrophomonas sp. was selected to enzymatically catalyze the saponification of Ax esters to produce free all‐trans‐Ax. Through single‐factor experiments and a Box–Behnken design, the optimal fermentation conditions were determined as follows: a seed culture age of 37.79 h, an inoculum concentration of 5.92%, and an initial broth pH of 6.80. Under these conditions, a fermentation curve was drawn, and the optimal fermentation time was shown to be 60 h. At 60 h, the degradation rate of the Ax esters was 98.08%, and the yield of free all‐trans‐Ax was 50.130 μg/mL. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:649–656, 2016     

ε-聚赖氨酸高产菌选育与发酵性能评价

【目的】选育ε-聚赖氨酸(ε-PL)高产菌,并探究不同碳源对其发酵性能的影响。【方法】借助基因组重排和核糖体工程两种育种手段强化ε-PL产生菌的合成能力,并利用p H冲击工艺评价不同碳源对ε-PL发酵的影响。【结果】经过4轮基因组重排和4轮核糖体工程连续选育,获得1株高产突变株Streptomyces albulus GS114,其摇瓶ε-PL产量达到3.0 g/L,较出发菌提高了1.7倍。该改造菌株在5 L发酵罐中分别以葡萄糖和甘油为碳源进行192 h的补料-分批发酵时,ε-PL发酵产量分别达到了43.4 g/L和45.7 g/L,较出发菌提高了11.0%和14.9%,而菌体量分别减少了24.0%和33.2%,ε-PL得率提高了34.2%和30.7%。【结论】基因组重排结合核糖体工程育种是一种有效的ε-PL高产菌选育手段,研究结果将为ε-PL高产菌改造和工业生产碳源选择提供直接指导。     

HAWAIIAN SKIRT regulates the quiescent center‐independent meristem activity in Arabidopsis roots

Root apical meristem (RAM) drives post‐embryonic root growth by constantly supplying cells through mitosis. It is composed of stem cells and their derivatives, the transit‐amplifying (TA) cells. Stem cell organization and its maintenance in the RAM are well characterized, however, their relationships with TA cells remain unclear. SHORTROOT (SHR) is critical for root development. It patterns cell types and promotes the post‐embryonic root growth. Defective root growth in the shr has been ascribed to the lack of quiescent center (QC), which maintains the surrounding stem cells. However, our recent investigation indicated that SHR maintains TA cells independently of QC by modulating PHABULOSA (PHB) through miRNA165/6. PHB controls TA cell activity by modulating cytokinin levels and type B Arabidopsis Response Regulator activity, in a dosage‐dependent manner. To further understand TA cell regulation, we conducted a shr suppressor screen. With an extensive mutagenesis screen followed by genome sequencing of a pooled F₂ population, we discovered two suppressor alleles with mutations in HAWAIIAN SKIRT (HWS). HWS, encoding an F‐box protein with kelch domain, is expressed, partly depending on SHR, in the root cap and in the pericycle of the differentiation zone. Interestingly, root growth in the shr hws was more active than the wild‐type roots for the first 7 days after germination, without recovering QC. Contrary to shr phb, shr hws did not show a recovery of cytokinin signaling. These indicate that HWS affects QC‐independent TA cell activities through a pathway distinctive from PHB.     

Purification,Chemical Characterization,and Bioactivity of an Extracellular Polysaccharide Produced by the Marine Sponge Endogenous Fungus <Emphasis Type="Italic">Alternaria</Emphasis> sp. SP-32

Marine sponges are ancient and simple multicellular filter-feeding invertebrates attached to solid substrates in benthic habitats and host a variety of fungi both inside and on their surface because of its unique ingestion and digest system. Investigation on marine sponge-associated fungi mainly focused on the small molecular metabolites, yet little attention had been paid to the extracellular polysaccharides. In this study, a homogeneous extracellular polysaccharide AS2-1 was obtained from the fermented broth of the marine sponge endogenous fungus Alternaria sp. SP-32 using ethanol precipitation, anion-exchange, and size-exclusion chromatography. Results of chemical and spectroscopic analyses showed that AS2-1 was composed of mannose, glucose, and galactose with a molar ratio of 1.00:0.67:0.35, and its molecular weight was 27.4 kDa. AS2-1 consists of a mannan core and a galactoglucan chain. The mannan core is composed of (1→6)-α-Manp substituted at C-2 by (1→2)-α-Manp with different degrees of polymerization. The galactoglucan chain consists of (1→6)-α-Glcp residues with (1→6)-β-Galf residues attached to the last glucopyranose residue at C-6. (1→6)-β-Galf residues have additional branches at C-2 consisting of disaccharide units of (1→2)-β-Galf and (1→2)-α-Glcp residues. The glucopyranose residue of the galactoglucan chain is linked to the mannan core. AS2-1 possessed a high antioxidant activity as evaluated by scavenging of 1,1-diphenyl-2-picrylhydrazyl and hydroxyl radicals in vitro. AS2-1 was also evaluated for cytotoxic activity on Hela, HL-60, and K562 cell lines by the MTT and SRB methods. The investigation demonstrated that AS2-1 was a novel extracellular polysaccharide with different characterization from extracellular polysaccharides produced by other marine microorganisms.