高山被孢霉Δ6Ⅱ-脱饱和酶的克隆、表达和功能鉴定

产油微生物高山被孢霉中Δ6脱饱和酶是决定ω3/ω6脂肪酸代谢流的关键酶,该酶对不同底物的催化特性直接决定该菌体内脂肪酸的流向。在已完成基因组测序的高山被孢霉ATCC32222中经注释发现它存在两种Δ6脱饱和酶(Δ6-Ⅰ和Δ6-Ⅱ),选择对其中的Δ6-Ⅱ脱饱和酶进行克隆、表达和功能鉴定。首先以p YES2/NT C质粒为骨架构建了Δ6-Ⅱ脱饱和酶基因(FADS6-Ⅱ)的表达载体(p YES2/NT C-FADS6-Ⅱ),并转化至酿酒酵母中进行诱导表达,进一步通过在重组菌培养基中添加Δ6脱饱和酶的底物来考察Δ6-Ⅱ脱饱和酶对各底物的偏好作用。实验结果表明,在分别添加0.5 mmol/L亚油酸(LA)和0.5 mmol/Lα-亚麻酸(ALA)时,Δ6-Ⅱ脱饱和酶对LA的转化率为42.94%,对ALA没有催化作用。而当底物添加方式改为同时添加LA和ALA时(分别为0.25 mmol/L),Δ6-Ⅱ脱饱和酶对LA的转化率为37.12%,对ALA仍没有催化作用。该实验结果为高山被孢霉中Δ6-Ⅱ脱饱和酶的催化功能研究提供了理论依据。     

Polyunsaturated fatty acids production by solid-state fermentation on polyurethane foam by Mortierella alpina

Polyunsaturated fatty acids (PUFAs) are essential in healthy diets and their production is extremely important. Natural sources of PUFAs includes animal and aquatic products such as marine fish oil, however there are several limitations such as the decrease of fish stocks throughout the world. Thus, microbial oils are a preferable source of PUFAs. Herein, it was studied the production of PUFAs by Mortierella alpina under solid-state fermentation (SSF) using polyurethane foam as inert substrate and synthetic medium or lignocellulosic hydrolysate as source of C, N, and other nutrients. Several parameters of fermentation conditions were evaluated as carbon source, inductors addition, ratio C/N and temperature. The highest amount of total PUFAs per mass of solid (535.41 ± 24.12 mg/g), linoleic acid (129.66 ± 5.84 mg/g), and α-linoleic acid (401.93 ± 18.10 mg/g) were produced when the culture medium contained 20 g/L glucose, 10% (w/v) linseed oil, the C/N ratio was adjusted to 25 and the incubation temperature was 25°C for 3 days decreasing to 16°C on the remaining 4 days of fermentation. In addition, a hemicellulosic hydrolysate can be used as low-cost substrate to produce PUFAs, although the production was lower than the achieved with synthetic medium. SSF showed an interesting technology for microbial PUFAs production.     

Age structure and age‐related differences in molt status and fuel deposition of Dunlins during the nonbreeding season at Chongming Dongtan in east China

ABSTRACT Although most shorebirds exhibit deferred migration and deferred breeding during their first summer, Dunlins (Calidris alpina) migrate to breeding areas and breed during their first summer. First‐year and adult Dunlins should, therefore, have similar fueling and molt patterns if energetic and physiological constraints are responsible for deferred migration. From 2006 to 2008, we examined the age structure of Dunlins during the nonbreeding season at Chongming Dongtan, an estuarine wetland in the Yangtze River estuary in east China, and examined the effects of date, age, and molt status on fuel deposition during migration and during the winter. The Dunlin population at Chongming Dongtan was composed primarily of first‐year birds. Most adults and first‐year birds arrived together in late August. Regression analyses indicated that age, date, and molt status affected fuel deposition (as indicated by body mass) of Dunlins. Adults had significantly greater fuel deposits than first‐year Dunlins near the end of northward migration (May: adults 70.8 ± 6.4 g, first‐year 63.8 ± 8.0 g) and at the start of southward migration (September: adults 50.2 ± 6.1 g, first‐year 47.2 ± 4.9 g). Adults also had significantly higher fuel deposition rates than first‐year Dunlins during northward migration. Nonetheless, first‐year Dunlins migrate and breed in their first summer. Thus, other factors, such as migration distance and body size, may be more important in determining if first‐year shorebirds defer migration during their first spring and summer. During boreal spring and autumn, first‐year Dunlins in active body molt had greater body mass than those that had not initiated body molt or those in suspended molt, and premigratory fuel deposits for northward migration were greatest after prealternate molt was completed. These results suggest that body molt requires additional fuel deposits and imposes a constraint on fuel deposition for migratory flights.