scholarly journals Assessment on the oil accumulation by knockdown of triacylglycerol lipase in the oleaginous diatom Fistulifera solaris

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yoshiaki Maeda ◽  
Kahori Watanabe ◽  
Marshila Kaha ◽  
Yusuke Yabu ◽  
Tomoko Yoshino ◽  
...  
Keyword(s):  
Wild Type Strain ◽  
Positive Impact ◽  
Lipid Production ◽  
Single Type ◽  
Oil Accumulation ◽  
Triacylglycerol Lipase ◽  
Lipid Degradation ◽  
Lipid Catabolism ◽  
Conventional Culture ◽  
Growth System

AbstractMicroalgae are promising producers of biofuel due to higher accumulation of triacylglycerol (TAG). However, further improvement of the lipid metabolism is critical for feasible application of microalgae in industrial production of biofuel. Suppression of lipid degradation pathways is a promising way to remarkably increase the lipid production in model diatoms. In this study, we established an antisense-based knockdown (KD) technique in the marine oleaginous diatom, Fistulifera solaris. This species has a capability to accumulate high content of lipids. Tgl1 KD showed positive impact on cell growth and lipid accumulation in conventional culture in f/2 medium, resulting in higher oil contents compared to wild type strain. However, these impacts of Tgl1 KD were slight when the cells were subjected to the two-stage growth system. The Tgl1 KD resulted in slight change of fatty acid composition; increasing in C14:0, C16:0 and C16:1, and decreasing in C20:5. This study indicates that, although Tgl1 played a certain role in lipid degradation in F. solaris, suppression of only a single type of TAG lipase was not significantly effective to improve the lipid production. Comprehensive understanding of the lipid catabolism in this microalga is essential to further improve the lipid production.

scholarly journals Improving the production of AHL lactonase AiiO-AIO6 from Ochrobactrum sp. M231 in intracellular protease-deficientBacillus subtilis

2020 ◽  
Author(s):  
Rui Xia ◽  
Yalin Yang ◽  
Xingliang Pan ◽  
Chenchen Gao ◽  
Yuanyuan Yao ◽  
...  
Keyword(s):  
Wild Type Strain ◽  
Positive Impact ◽  
Cell Communication ◽  
Quorum Quenching ◽  
Catalytic Triad ◽  
Heterologous Proteins ◽  
Extracellular Production ◽  
Expression Plasmid ◽  
Encoding Genes ◽  
Hydrolase Family

Abstract Quorum quenching (QQ) blocks bacterial cell-to-cell communication (i.e., quorum sensing), and is a promising antipathogenic strategy to control bacterial infection via inhibition of virulence factor expression and biofilm formation. QQ enzyme AiiO-AIO6 from Ochrobactrum sp. M231 has several excellent properties and shows biotherapeutic potential against important bacterial pathogens of aquatic species. AiiO-AIO6 can be secretory expressed in Bacillus subtilis via a non-classical secretion pathway.To improve AiiO-AIO6 production, four intracellular protease-deletion mutants of B. subtilis1A751 were constructed by individually knocking out the intracellular protease-encoding genes (tepA, ymfH, yrrNandywpE). The AiiO-AIO6 expression plasmid pWB-AIO6BS was transformed into the B. subtilis 1A751 and its four intracellular protease-deletion derivatives. Results showed that all recombinant intracellular protease-deletion derivatives (BSΔtepA, BSΔymfH, BSΔyrrNand BSΔywpE) had a positive impact on AiiO-AIO6 production. The highest amount of AiiO-AIO6 extracellular production of BSΔywpE in shake flask reached 1416.47 U/mL/OD600, which was about 121% higher than that of the wild-type strain. Furthermore, LC-MS/MS analysis of the degrading products of 3-oxo-C8-HSL by purification of AiiO-AIO6 indicated that AiiO-AIO6 was an AHL-lactonase which hydrolyzes the lactone ring of AHLs. Phylogenetic analysis showed that AiiO-AIO6was classified as a member of the α/β hydrolase family with a conserved “nucleophile-acid-histidine” catalytic triad. In summary, this study showed that intracellular proteases were responsible for the reduced yields of heterologous proteins and provided an efficient strategy to enhance the extracellular production of AHL lactonase AiiO-AIO6.

Peptides’ involvement in Pseudomonas putida biofilm formation

2020 ◽  
Author(s):  
Riho Teras ◽  
Hanna Ainelo ◽  
Marge Puhm
Keyword(s):  
Pseudomonas Putida ◽  
Mutant Strain ◽  
Type Strain ◽  
Deletion Mutant ◽  
Wild Type Strain ◽  
Positive Impact ◽  
Proteinase K ◽  
Wild Type ◽  
Positive Effect ◽  
Positively Charged

<p>Pseudomonas putida rapidly forms a biofilm, after which its biomass usually disperses to half its initial amount. We have observed different biofilm dynamics of P. putida in a complex medium LB and a minimal medium M9+glc+CAA and inquired about the importance of extracellular factors for the formation of P. putida biofilm.</p> <p>The proteinaceous component of LB increases the biomass of P. putida biofilm. Supplementation of M9 with tryptone but not CAA increased the biofilm biomass. Proteinase K treatment of LB medium reduced the biomass of P. putida biofilm. At the same time, growth rate or maximum OD of planktic bacteria in used media did not correlate with biofilm biomass of the same media. Thus, peptides appeared to have a positive effect on the biofilm as an extracellular factor and not as a source of C and N.</p> <p>We replaced tryptone in M9 medium with positively charged poly-L-lysine (MW. 1000-5000 Da), negatively charged poly-L-glutaminic acid (MW. 1500-5500 Da) or neutral poly-LD-alanine (MW. 3000-7000). Poly-lysine and poly-glutamic acid had a slight positive effect on the biomass of P. putida wild type strain PSm biofilm and poly-alanine did not affect the biofilm.</p> <p>We have previously shown that overexpression of fis in P. putida strain F15 increases biofilm biomass by increasing the lapA expression, the main adhesin gene of biofilm. Using media similar to that used for the wild-type strain for strain F15, we ascertained that only poly-lysine out of these three polypeptides restored the positive effect of fis-overexpression on the biofilm biomass. At the same time, the positive impact of fis-overexpression was absent in lapA deletion mutant strain, but not in lapF deletion mutant strain.</p> <p>In conclusion, the formation of P. putida biofilm depends on polypeptides in the environment. The enhancing effect of positively charged polypeptides appears to be evident in the presence of LapA, a key factor for P. putida biofilm.</p>

scholarly journals Increased Accumulation of Medium-Chain Fatty Acids by Dynamic Degradation of Long-Chain Fatty Acids in Mucor circinelloides

Genes ◽  
2020 ◽  
Vol 11 (8) ◽  
pp. 890
Author(s):  
Syed Ammar Hussain ◽  
Alexis Garcia ◽  
Md. Ahsanul Kabir Khan ◽  
Shaista Nosheen ◽  
Yao Zhang ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Wild Type Strain ◽  
Lipid Production ◽  
Mucor Circinelloides ◽  
Long Chain Fatty Acids ◽  
Wild Type ◽  
Medium Chain ◽  
Long Chain ◽  
Chain Fatty Acids

Concerns about global warming, fossil-fuel depletion, food security, and human health have promoted metabolic engineers to develop tools/strategies to overproduce microbial functional oils directly from renewable resources. Medium-chain fatty acids (MCFAs, C8–C12) have been shown to be important sources due to their diverse biotechnological importance, providing benefits ranging from functional lipids to uses in bio-fuel production. However, oleaginous microbes do not carry native pathways for the production of MCFAs, and therefore, diverse approaches have been adapted to compensate for the requirements of industrial demand. Mucor circinelloides is a promising organism for lipid production (15–36% cell dry weight; CDW) and the investigation of mechanisms of lipid accumulation; however, it mostly produces long-chain fatty acids (LCFAs). To address this challenge, we genetically modified strain M. circinelloides MU758, first by integrating heterologous acyl-ACP thioesterase (TE) into fatty acid synthase (FAS) complex and subsequently by modifying the β-oxidation pathway by disrupting the acyl-CoA oxidase (ACOX) and/or acyl-CoA thioesterase (ACOT) genes with a preference for medium-chain acyl-CoAs, to elevate the yield of MCFAs. The resultant mutant strains (M-1, M-2, and M-3, respectively) showed a significant increase in lipid production in comparison to the wild-type strain (WT). MCFAs in M-1 (47.45%) was sharply increased compared to the wild type strain (2.25%), and it was further increased in M-2 (60.09%) suggesting a negative role of ACOX in MCFAs production. However, MCFAs in M-3 were much decreased compared to M-1,suggesting a positive role of ACOT in MCFAs production. The M-2 strain showed maximum lipid productivity (~1800 milligram per liter per day or mg/L.d) and MCFAs productivity (~1100 mg/L.d). Taken together, this study elaborates on how the combination of two multidimensional approaches, TE gene over-expression and modification of the β-oxidation pathway via substantial knockout of specific ACOX gene, significantly increased the production of MCFAs. This synergistic approach ultimately offers a novel opportunity for synthetic/industrial biologists to increase the content of MCFAs.

scholarly journals Gene silencing of Sugar-dependent 1 (JcSDP1), encoding a patatin-domain triacylglycerol lipase, enhances seed oil accumulation in Jatropha curcas

2014 ◽  
Vol 7 (1) ◽  
pp. 36 ◽  
Author(s):  
Mi Kim ◽  
Seong Yang ◽  
Hui-Zhu Mao ◽  
Sivaramakrishnan P Veena ◽  
Jun-Lin Yin ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Jatropha Curcas ◽  
Seed Oil ◽  
Oil Accumulation ◽  
Triacylglycerol Lipase

scholarly journals Implementasi Cara Hidup Jemaat Mula-mula dalam Kisah Para Rasul 2: 41-47 bagi Pertumbuhan Gereja Masa Kini

2020 ◽  
Vol 2 (2) ◽  
pp. 127-140
Author(s):  
Andreas Sese Sunarko
Keyword(s):  
Church Growth ◽  
Positive Impact ◽  
Church Members ◽  
Way Of Life ◽  
Senior Pastor ◽  
The People ◽  
Starting Point ◽  
Daily Behavior ◽  
Growth System ◽  
Growth Method

The growth of church has been one of crucial issues that becomes a concentration point to church leaders, which is priestas the senior pastor or church council as the representative of local church members. The growth of church perceived as one of paramaters stating the existence of church to the people who are served as well as to other churches as ministry partner. The growth of church can be seen from two aspect is the quality of church members in terms of spiritual maturity reflected from the faith to God they whorship and in term of their daily behavior bringing positive impact. The other aspect is the quantity of church members from time to time.   The growth of church can  be achieved by the method or serving system that is believed by priest or church council to be implemented. In academic terminology, that method is also known as church growth system or method . They are many system or method of church growth . One of them is Cell Church system that is a system of naturaly growing church growth method with the starting point from the way of life the eraly church taken from Act 2:41-47. This idea, later on, can be implemented by today’s churches is improving church growth.AbstrakPertumbuhan gereja menjadi salah satu isu penting yang terus menjadi konsentrasi bagi para pimpinan gereja, baik itu Pendeta sebagai Gembala Jemaat atau Majelis sebagai representasi jemaat lokal. Pertumbuhan gereja dinilai sebagai salah satu parameter eksistensi gereja tersebut dimata jemaat yang dilayaninya maupun dimata gereja-gereja lain sebagai mitra pelayanan. Pertumbuhan gereja bisa dilihat dari dua segi yaitu segi kualitas yang ditandai dengan kematangan jemaat atas nilai-nilai relegiositasnya yang terpancar dari tingkat keyakinannya terhadap pribadi Allah yang disembahnya dan terpancar dari perilaku sehari-harinya yang mendatangkan dampak positif. Adapun segi yang lain adalah segi kuantitas yang ditandai dengan pertumbuhan jemaat yang terlihat dari bertambahnya jumlah jemaat dari waktu ke waktu. Pertumbuhan gereja ini dapat terwujud dengan metode dan sistem pelayanan yang diyakini oleh Pendeta atau Majelis untuk diterapkan atau dalam dunia akademis disebut dengan metode atau sistem pertumbuhan gereja. Ada banyak metode atau sistem pertumbuhan gereja yang ada diantaranya metode atau sistem Gereja Sel, metode atau sistem pertumbuhan Gereja Modern dan lain sebagainya. Melalui tulisan ini penulis ingin menunjukan salah satu metode pertumbuhan gereja yang bertitik tolak pada cara hidup jemaat yang pertama sebagaimana dicatat dalam Kisah Para Rasul 2 : 41-47 yang nantinya dapat diterapkan oleh gereja-gereja masa kini dalam melakukan proses pertumbuhan gereja.

scholarly journals Biochemical Modulation of Lipid Pathway in MicroalgaeDunaliellasp. for Biodiesel Production

2015 ◽  
Vol 2015 ◽  
pp. 1-12 ◽  
Author(s):  
Ahmad Farhad Talebi ◽  
Masoud Tohidfar ◽  
Seyedeh Mahsa Mousavi Derazmahalleh ◽  
Alawi Sulaiman ◽  
Azhari Samsu Baharuddin ◽  
...  
Keyword(s):  
Feedback Inhibition ◽  
Lipid Production ◽  
Cetane Number ◽  
Nile Red ◽  
Transcript Abundance ◽  
Point Of View ◽  
Biodiesel Production ◽  
Biochemical Modulation ◽  
Oil Accumulation ◽  
Microalgal Biodiesel

Exploitation of renewable sources of energy such as algal biodiesel could turn energy supplies problem around. Studies on a locally isolated strain ofDunaliellasp. showed that the mean lipid content in cultures enriched by 200 mg L−1myoinositol was raised by around 33% (1.5 times higher than the control). Similarly, higher lipid productivity values were achieved in cultures treated by 100 and 200 mg L−1myoinositol. Fluorometry analyses (microplate fluorescence and flow cytometry) revealed increased oil accumulation in the Nile red-stained algal samples. Moreover, it was predicted that biodiesel produced from myoinositol-treated cells possessed improved oxidative stability, cetane number, and cloud point values. From the genomic point of view, real-time analyses revealed that myoinositol negatively influenced transcript abundance ofAccDgene (one of the key genes involved in lipid production pathway) due to feedback inhibition and that its positive effect must have been exerted through other genes. The findings of the current research are not to interprete that myoinositol supplementation could answer all the challenges faced in microalgal biodiesel production but instead to show that “there is a there there” for biochemical modulation strategies, which we achieved, increased algal oil quantity and enhanced resultant biodiesel quality.

scholarly journals Hypericin-mediated sonodynamic therapy induces autophagy and decreases lipids in THP-1 macrophage by promoting ROS-dependent nuclear translocation of TFEB

2016 ◽  
Vol 7 (12) ◽  
pp. e2527-e2527 ◽  
Author(s):  
Xuesong Li ◽  
Xin Zhang ◽  
Longbin Zheng ◽  
Jiayuan Kou ◽  
Zhaoyu Zhong ◽  
...  
Keyword(s):  
Nuclear Translocation ◽  
Lipid Uptake ◽  
Sonodynamic Therapy ◽  
Lipid Degradation ◽  
Lipid Catabolism ◽  
Lipid Efflux ◽  
Abca1 Expression ◽  
Transcription Factor Eb ◽  
Ros Scavenger ◽  
Cholesterol Transporters

Abstract Lipid catabolism disorder is the primary cause of atherosclerosis. Transcription factor EB (TFEB) prevents atherosclerosis by activating macrophage autophagy to promote lipid degradation. Hypericin-mediated sonodynamic therapy (HY-SDT) has been proved non-invasively inducing THP-1-derived macrophage apoptosis; however, it is unknown whether macrophage autophagy could be triggered by HY-SDT to influence cellular lipid catabolism via regulating TFEB. Here, we report that HY-SDT resulted in the time-dependent THP-1-derived macrophage autophagy activation through AMPK/AKT/mTOR pathway. Besides, TFEB nuclear translocation in macrophage was triggered by HY-SDT to promote autophagy activation and lysosome regeneration which enhanced lipid degradation in response to atherogenic lipid stressors. Moreover, following HY-SDT, the ABCA1 expression level was increased to promote lipid efflux in macrophage, and the expression levels of CD36 and SR-A were decreased to inhibit lipid uptake, both of which were prevented by TFEB knockdown. These results indicated that TFEB nuclear translocation activated by HY-SDT was not only the key regulator of autophagy activation and lysosome regeneration in macrophage to promote lipolysis, but also had a crucial role in reverse cholesterol transporters to decrease lipid uptake and increase lipid efflux. Reactive oxygen species (ROS) were adequately generated in macrophage by HY-SDT. Further, ROS scavenger N-acetyl-l-cysteine abolished HY-SDT-induced TFEB nuclear translocation and autophagy activation, implying that ROS were the primary upstream factors responsible for these effects during HY-SDT. In summary, our data indicate that HY-SDT decreases lipid content in macrophage by promoting ROS-dependent nuclear translocation of TFEB to influence consequent autophagy activation and cholesterol transporters. Thus, HY-SDT may be beneficial for atherosclerosis via TFEB regulation to ameliorate lipid overload in atherosclerotic plaques.

FAX2 Mediates Fatty Acid Export from Plastids in Developing Arabidopsis Seeds

2019 ◽  
Vol 60 (10) ◽  
pp. 2231-2242 ◽  
Author(s):  
Yinshuai Tian ◽  
Xueyan Lv ◽  
Guilan Xie ◽  
Linghui Wang ◽  
Tingwei Dai ◽  
...  
Keyword(s):  
Target Gene ◽  
Seed Oil ◽  
Mutant Cell ◽  
Lipid Production ◽  
Building Blocks ◽  
Wild Type ◽  
Oil Accumulation ◽  
Oil Synthesis ◽  
Family Protein ◽  
Synthesis Activity

Abstract Vegetable oils are mainly stored in the form of triacylglycerol (TAG) in oilseeds. Fatty acids (FAs), one of the building blocks for TAG assembly, are synthesized in plastids and then exported to the endoplasmic reticulum for storage oil synthesis. A recent study demonstrated that the export of FAs from plastids was mediated by a FAX (FA export) family protein. However, the significance of FAs export from plastid during seed oil accumulation has not been investigated. In this study, we found that FAX2 was highly expressed in developing Arabidopsis seeds and the expression level was consistent with FAs synthesis activity. FAX2 mutant seeds showed an approximately 18% reduction of lipid levels compared with wild-type seeds. By contrast, overexpression of FAX2 enhanced seed lipid accumulation by up to 30%. The FAs export activity of FAX2 was confirmed by yeast mutant cell complementation analysis. Our results showed that FAX2 could interact with other proteins to facilitate FAs transport. Taken together, these results indicate that FAX2-mediated FA export from plastids is important for seed oil accumulation, and that FAX2 can be used as a target gene for increasing lipid production in oilseeds.

Hope, exploration, and equilibrated action schemes

2019 ◽  
Vol 42 ◽  
Author(s):  
Davood G. Gozli ◽  
Ci Jun Gao
Keyword(s):  
Single Type ◽  
Reward Seeking ◽  
Stable Scheme

AbstractThe concepts want, hope, and exploration cannot be organized in relation to a single type of motive (e.g., motive for food). They require, in addition, the motive for acquiring and maintaining a stable scheme that enables reward-directed activity. Facing unpredictability, the animal has to seek not only reward, but also a new equilibrated state within which reward seeking is possible.

Multidisciplinary team decision is rare and decreasing in percutaneous vascular interventions despite positive impact on in-hospital outcomes

VASA ◽  
2019 ◽  
Vol 48 (3) ◽  
pp. 262-269 ◽  
Author(s):  
Christian-Alexander Behrendt ◽  
Tilo Kölbel ◽  
Thea Schwaneberg ◽  
Holger Diener ◽  
Ralf Hohnhold ◽  
...  
Keyword(s):  
Multidisciplinary Team ◽  
Positive Impact ◽  
Cross Sectional Study ◽  
Invasive Treatment ◽  
Cross Sectional ◽  
Hospital Outcomes ◽  
Vascular Intervention ◽  
Medical Specialties ◽  
Team Decision ◽  
The Impact

Abstract. Background: Worldwide prevalence of peripheral artery disease (PAD) is increasing and peripheral vascular intervention (PVI) has become the primary invasive treatment. There is evidence that multidisciplinary team decision-making (MTD) has an impact on in-hospital outcomes. This study aims to depict practice patterns and time changes regarding MTD of different medical specialties. Methods: This is a retrospective cross-sectional study design. 20,748 invasive, percutaneous PVI of PAD conducted in the metropolitan area of Hamburg (Germany) were consecutively collected between January 2004 and December 2014. Results: MTD prior to PVI was associated with lower odds of early unsuccessful termination of the procedures (Odds Ratio 0.662, p < 0.001). The proportion of MTD decreased over the study period (30.9 % until 2009 vs. 16.6 % from 2010, p < 0.001) while rates of critical limb-threatening ischemia (34.5 % vs. 42.1 %), patients´ age (70 vs. 72 years), PVI below-the-knee (BTK) (13.2 % vs. 22.4 %), and rates of severe TASC C/D lesions BTK (43.2 % vs. 54.2 %) increased (all p < 0.001). Utilization of MTD was different between medical specialties with lowest frequency in procedures performed by internists when compared to other medical specialties (7.1 % vs. 25.7 %, p < 0.001). Conclusions: MTD prior to PVI is associated with technical success of the procedure. Nonetheless, rates of MTD prior to PVI are decreasing during the study period. Future studies should address the impact of multidisciplinary vascular teams on long-term outcomes.

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