Biotechnology ofα-linolenic acid in oilseed rape(Brassica napus)using FAD2 and FAD3 from chia(Salvia hispanica)

α-Linolenic acid(ALA,18:3Δ9,12,15)is an essential fatty acid for humans since it is the precursor for the biosynthesis of omega-3 long-chain polyunsaturated fatty acids(LC-PUFA).Modern people generally suffer from deficiency of ALA because most staple food oils are low or lack ALA content.Biotechnological enrichment of ALA in staple oil crops is a promising strategy.Chia(Salvia hispanica)has the highest ALA content in its seed oil among known oil crops.In this study,the FAD2 and FAD3 genes from chia were engineered into a staple oil crop,oilseed rape(Brassica napus),via Agrobaterium tumefaciens-mediated transformation of their LP4-2A fusion gene construct driven by the seed-specific promoter P_(NapA).In seeds of T0,T1,and T2 lines,the average ALA contents were 20.86,23.54,and 24.92%,respectively,which were 2.21,2.68,and 3.03 folds of the non-transformed controls(9.42,8.78,and 8.22%),respectively.The highest seed ALA levels of T0,T1,and T2 plants were 38.41,35.98,and 39.19%respectively,which were 4.10-4.77 folds of the respective controls.FA-pathway enzyme genes(BnACCD,BnFATA,BnSAD,BnSCD,BnDGAT1,BnDGAT2,and BnDGAT3)and positive regulatory genes(BnWRI1,BnLEC1,BnL1L,BnLEC2,BnABI3,BnbZIP67,and BnMYB96)were all significantly up-regulated.In contrast,BnTT1,BnTT2,BnTT8,BnTT16,BnTTG1,and BnTTG2,encoding negative oil accumulation regulators but positive secondary metabolism regulators,were all significantly down-regulated.This means the foreign ShFAD2-ShFAD3 fusion gene,directly and indirectly,remodeled both positive and negative loci of the whole FA-related network in transgenic B.napus seeds.

Formulation ofα-linolenic acid microemulsion free of co-surfactant

A novel class ofα-linolenic acid-in-water microemulsion free of co-surfactant was investigated as potential food delivery systems.Rough demarcation within the transparent region was deduced from the results of conductivity and polarizing optical microscopy.The microemulsion mean hydrodynamic diameter and characterization were determined by dynamic light scattering and negative-staining TEM.The location of ALA molecules in the microemulsion formulations was determined by ~1H NMR spectroscopy.

α-Linolenic acid alleviates aluminium chloride-induced toxicity in PC12 cells by activation of PKA-CREB-BDNF signaling pathway

Aluminum has been associated with neurodegenerative diseases.ALA(α-linolenic acid),an essential dietary component for human health,possesses prominent biological activities.Herein,we aim to explore the neuroprotective effects of ALA on aluminum toxicity and reveal the underlying mechanism.Results show that aluminum chloride(denoted as Al)enabled cell viability decline and apoptosis with oxidative stress and mitochondrial damage in differentiated rat pheochromocytoma cells(PC12)for 24 h incubation.Compared with Al(10 mmol/L)treatment alone,ALA(50μmol/L)pretreatment for 24 h significantly enhanced cell viability by 28.40%,and hindered cell apoptosis by 12.35%,together with recovering redox state balance and alleviating mitochondrial damage.It was measured that ALA treatment upregulated Bcl-2 expression and down-regulated Bax level,accompanied with an expression decline of caspase-3 and caspase-9.Meanwhile,ALA pretreatment was proved to increase protein kinase A(PKA)expression and to promote phosphorylation of cAMP response element-binding protein(p-CREB),resulting in elevation on the level of brain-derived neurotrophic factor(BDNF).The above results showed that ALA attenuated Al toxicity in PC12 cells by mediating the PKA-CREBBDNF signaling pathway.

深黄被孢霉在不同培养条件下合成GLA和代谢副产物脂肪酸的分析

本文研究了葡萄糖、铵离子、金属离子、缓冲剂、表面活性剂和温度等因素对深黄被孢霉合成GLA的影响。并将ΣC18/ΣC16、IUFA、(L+G)/O和G/L等值作为脂肪酸分析的指标来考察GLA合成量改变的机理。结果表明:铵离子对GLA合成中脂肪酸△6-脱氢、不饱和度和碳链延长有明显的影响;Ca2+对脂肪酸△12-脱氢和不饱和程度有促进作用;0.3%NaAC可对△6-和△12-脂肪酸脱氢酶活性有明显的增强作用;低温培养可明显增强△6-脂肪酸脱氢和碳链延长的作用。

Feeding Calcium Salts of Linseed Oil on Metabolic Status and Reproductive Traits in Grazing Dairy Cows

This experiment aimed to evaluate the effect of calcium salts of linseed oil (rich in alpha-linolenic acid (ALA, C18:3n-3) on metabolic and reproductive traits in high-producing dairy cows under grazing. Thirty-six Holstein dairy cows were randomly assigned, in a complete block design, to receive ALA supplementation (0.85 kg∙day−1 of calcium salts of linseed oil) or to remain as untreated control (CON). The concentrate was formulated to offer the same amount of energy across treatments (CON cows received an extra kg of corn to compensate for the higher energy density of ALA treatment). A PMR + Alfalfa pasture was offered to all cows at the same time. A fixed time artificial insemination (FTAI) at 80 DIM, preceded by a Presynch plus Ovsynch protocol was implemented for the first service and later, on return to estrus, heat detection and artificial insemination (AI) were performed. Pregnancy diagnosis was checked at 30, 42, 60, and 90 d after AI. Blood and milk samples were taken biweekly. Treatment affected plasma cholesterol concentration (160.36 vs. 186.70 mg∙dl−1, p = 0.03, for ALA and CON, respectively) and on size of corpus luteum (CL, 17.6 vs. 13.7 mm, p = 0.02, for ALA and CON, respectively). Supplementation tended (p = 0.136) to increase conception rate by 200 DIM (81.69% vs. 55.43% in ALA and CON cows, respectively). However, treatment had no effect (p > 0.05) on body weight (BW), body condition score (BCS), and circulating levels of beta-hydroxybutyrate (BHBA), glucose, insulin, growth hormone (GH) and insulin-like growth factor (IGF-I). Our results suggest that supplementation with calcium salts of linseed oil could enhance ovarian function without affecting energy metabolism in early lactation dairy cows.

The Role of Jasmonates as Antibulbing Substances in the Bulb Formation of Onion

Onion plants form spherical bulbs under long-day conditions.Substances regulating bulb formation remain unknown.In the course of chemical studies on the bulb formation,α-linolenic acid was isolated from onion extracts as an antibulbing substance,the amount of which was synchronized with the bulb formation.Since allene oxide synthase inhibitor canceled the antibulbing activity ofα-linolenic acid,it was disclosed that jasmonic acid concerns this regulation.Structure-activity-relationship study revealed that its(3R,7S)stereochemistry is necessary for showing its antibulbing activity.It is concluded that(3R,7S)-jasmonate derived fromα-linolenic acid actually participates in the regulation of bulb formation.

被孢霉产γ-亚麻酸(GLA)后处理工艺的研究进展

综述了被孢霉产γ-亚麻酸(GLA)后处理工艺中干菌体的获得、菌体中油脂的提取、GLA的富集及GLA含量的测定,拟得发酵后理想的产物及准确检测GLA含量的后处理方法。

利用真菌生产GLA、Ara和EPA的研究进展

综述了GLA、Ara、EPA的生理作用、天然来源和利用真菌生产的研究进展。研究发现真菌资源特别是低等真菌是除动、植物外GLA、Ara和EPA的另一个重要来源 ,通过菌种选育可以得到与动、植物产量相近的高产菌株 ,并在低温培养时有利于多不饱和脂肪酸的形成 ,细胞内微粒体膜的苹果酸酶活性与菌体油脂的多不饱和脂肪酸含量存在一定的正相关关系。此外 ,利用微生物转化技术 ,以天然油脂为底物来生产GLA。

Evaluation of anti-tubercular activity of linolenic acid and conjugatedlinoleic acid as effective inhibitors against Mycobacterium tuberculosis

Objective: To evaluate a new pharmacological activity/effect of linolenic acid(α- and γ-form) and conjugated-linoleic acid(CLA) causing antibacterial activity against Mycobacterium tuberculosis(Mtb). Methods: The anti-Mtb activity/effect of linolenic acid and CLA were determined using different anti-Mtb indicator methods such as resazurin microtiter assay(REMA) and MGIT 960 system assay. The Mtb was incubated with various concentrations(12.5–200) μg/m L of the compounds and anti-Mtb first-line drugs for 5 d in the REMA, and for 3 wk in MGIT 960 system assay. Results: Linolenic acid and CLA obviously indicated their anti-Mtb activity/effect by strongly inhibiting the growth/proliferation of Mtb in a dosedependent manner in the REMA and the MGIT 960 system assay. Interestingly, linolenic acid and CLA consistently induced anti-Mtb activity/effect by effectively inhibiting the growth/proliferation of Mtb in MGIT 960 system for 21 d with a single-treatment, and their minimum inhibitory concentrations were measured as 200 μg/m L respectively. Conclusions: These results demonstrate that linolenic acid and CLA not only have effective anti-Mtb activity/properties, but also induce the selective-anti-Mtb effects by strongly inhibiting and blocking the growth/proliferation of Mtb through a new pharmacological activity/action. Therefore, this study provides novel perspectives for the effective use of them and the potential that can be used as potent anti-Mtb candidate drugs, as well as suggests the advantage of reducing the cost and/or time for developing a new/substantive drug by effectively repurposing the existing drugs or compounds as one of new strategies for the global challenge of tuberculosis.

Fatty acid analysis in the seeds of 50 Paeonia ostii individuals from the same population

Tree peony seeds are rich in α-linolenic acid(ALA), and the peony seed oil is now being produced in China. Paeonia ostii is the most widely used tree peony species for oil extraction, which is commercially called Fengdan and treated as a single cultivar. Here, 50 P. ostii individuals from the same population in northern China were randomly selected for fatty acids(FAs) analysis. Thirteen FAs were isolated, and the most abundant five were palmitic acid(5.31–6.99%), stearic acid(1.22–2.76%), oleic acid(18.78–28.15%), linoleic acid(11.86–26.10%), and ALA(41.11–57.51%). There were significant individual differences of plants in FA quality and quantity and the linoleic acid content in Plant No. 48 even exceeded the scope of 1–99%. Further statistical analysis indicated that most of the individual FAs, saturated FAs, unsaturated FAs, and total FAs levels showed significant positive correlations to each other, whereas the seed yield per plant was independent and not correlated to the factors mentioned above. Ward’s hierarchical clustering results grouped the 50 plants into four clusters based on FA contents and seed yield, and the seven plants in Cluster IV were identified as good candidates for oil production. Our results confirmed that the individual differences did occur in P. ostii and Fengdan cannot be simply treated as one uniform cultivar. Also, these results may help simplify the selection of plants for oil peony breeding and accelerate the development of the oil peony industry.

Feeding Calcium Salts of Linseed Oil on Productive Performance and Milk Fatty Acid Profile in Grazing Dairy Cows

The objective of this study was to determine the effect of supplying calcium salts of linseed oil (Ca-FA) rich in omega-3 (α-linolenic acid) on the production and chemical composition of milk and its nutraceutical value in dairy cows in early lactation. The trial lasted 12 weeks (2 weeks for adaptation to lipids and 10 weeks of data collection). A total of 36 Holstein dairy cows with 58.0 ± 17.0 days in milk (DIM), 594.1 ± 92.4 kg BW, 2.6 ± 1.5 parity and 38.9 ± 9.3 kg milk day-1 were used in a randomized complete block design. The treatments were: 1) Omega-3 (O3): 5.2 kg DM day-1 of concentrate including 0.7 kg DM of Ca-FA + 13.5 kg DM day-1 of partial mixed ration (PMR) + 12 kg DM day-1 of alfalfa pasture (Medicago sativa) and 2) Control (C): diet similar to O3 but lipid supplementation was replaced by cracked corn grain so that the diets were isoenergetic. No treatment effect was detected (P > 0.05) for any milk production and composition variables, except for urea in milk that was slightly higher in O3 (P = 0.02). The treatment × week interaction was significant (P < 0.05) for fat yield and content, with differences (P < 0.01) only in the 3rd week of the data collection period in favor of group C (1.39 vs. 1.13 kg·day-1 and 3.86% vs. 3.23% for fat yield and content, respectively). Total DMI and PMR were similar (P > 0.05) between treatments. Concentrate intake was higher (P < 0.01) in C compared with O3. Pasture DMI tended (P = 0.06) to be greater for cows that received the O3 treatment compared with C. Total metabolizable energy (ME) intake was similar (P = 0.44) between treatments. No treatment effect was detected (P > 0.05) in rumen environment parameters. Supplementation with Ca-FA reduced (P < 0.05) the hypercholesterolemic fraction of milk (C12:0, C14:0 and C16:0, -13.6%, -7.4% and -9.0%, respectively). The concentration of α-linolenic acid (C18:3n-3) increased (108%, P < 0.01) in O3 group compared with group C. The absence of negative effects of lipids on the fat content of milk and ruminal fermentation suggests that protection by saponification was effective. The supplementation with Ca-FA (0.85 kg·day-1) improved the healthy value of the milk.

中医健脾单元疗法对强直性脊柱炎患者疗效及BGP、TRACP的影响

目的探讨中医健脾单元疗法对强直性脊柱炎(Ankylosing Spondylitis,AS)患者疗效及对血清骨钙素(bone gla protein,BGP)、抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRACP)的影响。方法按随机数字表将60例强直性脊柱炎患者分成治疗组(34例)和对照组(26例),治疗组采用中医健脾单元疗法,即新风胶囊+中医辨证论治+中药熏蒸治疗,对照组采用柳氮磺嘧啶(SASP)+中医辨证论治+中药熏蒸治疗。比较两组患者疗效、治疗前后生活质量、症状体征、急性时相反应物指标及对血清BGP、TRACP等指标变化。结果①中医证候疗效评价结果示两组总有效率与显效率比较,差异有统计学意义(P<0.05)。②两组患者治疗后生活质量总积分、焦虑自评量表(SAS)、抑郁自评量表(SDS)、疼痛严重性评估(VAS)、疾病活动指数(BASDAI)、躯体功能指数BASFI)、整体指数(BAS-G)、中医证候积分、超敏C反应蛋白(hs-CRP)、α酸性糖蛋白(α-AGP)与治疗前比较,差异均有统计学意义(P<0.01);治疗组治疗后血沉(ESR)与治疗前比较,差异有统计学意义(P<0.01);治疗后组间比较,两组生活质量总积分、SAS、SDS、VAS、BASDAI、BASFI、BAS-G、中医证候积分差异均有统计学意义(P<0.01)。③治疗后两组患者血清BGP及TRACP与治疗前比较,差异均有统计学意义(P<0.01),治疗后组间比较,差异亦有统计学意义(P<0.01)。结论中医健脾单元疗法能够提高AS患者的生活质量,改善临床症状体征、焦虑抑郁情绪及实验室指标,疗效优于对照组,其机制可能是通过上调BGP、下调TRACP,改善骨代谢水平,降低组织炎症反应,从而改善全身症状。

γ-亚麻酸高产菌株的选育及发酵产物的分离提取

以深黄被孢霉AS3.3410为出发菌株发醇产生γ-亚麻酸,其菌体得率为10%,油脂含量为27%,γ-亚麻酸含量为3.3%。经过紫外线诱变处理。得到变异株M_6,其菌体得率为25%,油脂含量为32.8%,γ-亚麻酸含量为8.84%。传代实验表明,M_6具有良好的遗传稳定性。经摇瓶发酵条件试验,选出了最佳培养基配方,以10升罐进行发酵,结果为:菌体得率29.3%,油脂含量44.7%,γ-亚麻酸含量9.44%。菌体油脂提取方法的研究表明,以乙醇和正己烷对湿菌体进行分步抽提效果较好。

γ-亚麻酸及其生理生化功能研究进展

γ-亚麻酸是一种具有多种生物学活性的多不饱和脂肪酸。多年研究表明:γ-亚麻酸存在于多种草本植物、藻类和低等真菌中,可通过多种生物学机制发挥降血脂、杀菌、抗炎、抗高血压、抗动脉粥样硬化等一系列生理生化功能。综述了γ-亚麻酸生物资源、代谢、生理生化功能及相关机制的研究进展。

肾性骨病80例临床分析

报告80例肾性骨病和钙、磷代谢变化的关系。其中男41例,女39例。发病年龄8～71岁,平均44岁。平均病程3.14±1.43年(1～10年)。骨质变化以骨质疏松居多占38例,骨软化和/或肾性佝偻病12例,纤维囊性骨炎10例,骨硬化10例。10例发生骨折。血钙平均值低于正常,血磷升高。24小时尿磷及尿钙减少。血清甲状旁腺激素(PTH)、降钙素(CT)、骨钙素(BGP),酸性磷酸酶(ACP)、抗酒石酸酸性磷酸酶(TrACP)及碱性磷酸酶(AKP)升高,均有显著性差异。

大麻籽油的特性及研究进展

大麻籽含有约25%-35%的油脂,20%-25%的蛋白质,20%-30%的碳水化合物,以及丰富的微量元素等。大麻籽油中不饱和脂肪酸的含量约为90%,其中必需脂肪酸的含量约80%。ω-6与ω-3多不饱和脂肪酸的比例约为3∶1,是理想的人体脂肪酸摄入比例,并且还含有γ-亚麻酸以及丰富的生育酚和植物甾醇。研究表明,大麻籽油在降低胆固醇、抗氧化、清除人体内自由基等方面具有显著的作用,并且是安全无毒的,是一种具有很高利用价值的功能性油脂。

添加植物油对发酵生产γ-亚麻酸的影响

以本实验室收藏的一株被孢霉突变株Ｍ９发酵生产γ－亚麻酸，在培养基中分别添加大豆油、花生油和橄榄油，可以使菌体得率和γ－亚麻酸的产量均有所提高。实验表明：非离子表面活性剂的加入可加强这种效果，植物油的添加在菌体生长的平衡期之前效果明显。

真菌发酵生产γ-亚麻酸的研究进展

综述了国内外微生物发酵生产γ -亚麻酸 (GLA)的研究进展 ,对菌株诱变选育研究和基因工程菌株的构建情况进行了阐述 ,并展望了其发展前景。

培养条件对钝顶螺旋藻(Sp)NS-90020脂肪酸组成和含量的影响

研究了不同培养条件对钝顶螺旋藻（Sp）NS－90020脂肪酸合成的影响。随着温度升高，其不饱和脂肪酸，γ一亚麻酸（GLA）相对含量降低，总脂肪酸含量升高，当温度为40℃时总脂肪酸和γ-亚麻酸绝对含量都达到最大值，分别为73．4mg／g干重和11．9mg／g干重；当培养基中NaCl浓度高于0．017mol／L时，其GLA相对含量降低，但低于0．0017mol／L时，对其脂肪酸组成无显著影响；氨水使其脂肪酸和GLA绝对含量升高，并在50mgN（NH3·H2O）时达到最大值，分别为67．96mg／g干重和13．63mg／g千重；暗处理92h使其总脂肪酸和GLA绝对含量升高；缺乏Fe2+或Mg2+或Mo2+时，其总脂肪酸和GLA绝对含量降低，而缺乏PO43-时，其总脂肪酸和GLA绝对含量略有升高。

功能性γ-亚麻酸代谢研究的进展

本文综述γ-亚麻酸的生理功能和代谢途径中△6-脂肪酸脱氢酶相关研究的进展,并提出可采用代谢工程的原理和方法对γ-亚麻酸做进一步深入探讨的观点。