1.Enhancement of harvesting efficiency and polyunsaturated fatty acid-rich lipid production of Aurantiochytrium sp. SW1 by co-cultivation with oleaginous fungus Cunninghamella bainieri 2A1
Nur Athirah Ahmad Jailani ; Pranesha Prabakaran ; Mohamed Yusuf Mohamed Nazir ; Wan Syaidatul Aqma Mohd Noor ; Aidil Abdul Hamid
Malaysian Journal of Microbiology 2022;18(6):620-628
Aims:
Thraustochytrids have been shown to be excellent lipid producers due to their ability to accumulate over 50% lipid (g/g biomass) containing up to 50% docosahexaenoic acid (DHA). However, efficient and cost-effective cell recovery of lipid-rich biomass has become a significant challenge at the industrial scale. In this study, we attempted to enhance the harvesting efficiency (HE) and the DHA content of Aurantiochytrium sp. through co-cultivation with a γ-linolenic acid (GLA)-producing oleaginous filamentous fungus, Cunninghamella bainieri 2A1.
Methodology and results:
A 72 h old C. bainieri 2A1 culture in the form of loose mycelia or pellets of various sizes was added into 72 h old Aurantiochytrium sp. cultures and further incubated for 48 h. The HE of Aurantiochytrium sp. was then determined by comparing the remaining OD values of the supernatant with and without minimal centrifugation at 4000× g. Results showed that 63.23% of HE was achieved without centrifugation from co-cultivation with dispersed mycelia. Higher HE between 96.71-99.55% was achieved when centrifugation was implemented, with the highest value resulting from co-cultivation with dispersed mycelia. These are higher than HE of centrifuged control cultures (80%) consisting of Aurantiochytrium sp. monocultures, suggesting that co-cultivation with C. bainieri 2A1 facilitates the recovery of Aurantiochytrium sp. cells. Moreover, the co-cultivation also resulted in a 28% increase in DHA compared to non-optimized cultures.
Conclusion, significance and impact of study
This study provides the first evidence of enhancement in harvesting and DHA content of oleaginous thraustochytrids that could be achieved through co-cultivation with oleaginous fungi.
Heterotrophic Processes
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Cunninghamella
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Eukaryota
2.Introns: The Functional Benefits of Introns in Genomes.
Genomics & Informatics 2015;13(4):112-118
The intron has been a big biological mystery since it was first discovered in several aspects. First, all of the completely sequenced eukaryotes harbor introns in the genomic structure, whereas no prokaryotes identified so far carry introns. Second, the amount of total introns varies in different species. Third, the length and number of introns vary in different genes, even within the same species genome. Fourth, all introns are copied into RNAs by transcription and DNAs by replication processes, but intron sequences do not participate in protein-coding sequences. The existence of introns in the genome should be a burden to some cells, because cells have to consume a great deal of energy to copy and excise them exactly at the correct positions with the help of complicated spliceosomal machineries. The existence throughout the long evolutionary history is explained, only if selective advantages of carrying introns are assumed to be given to cells to overcome the negative effect of introns. In that regard, we summarize previous research about the functional roles or benefits of introns. Additionally, several other studies strongly suggesting that introns should not be junk will be introduced.
DNA
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Eukaryota
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Genome*
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Introns*
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RNA
3.Fungal luminescence pathways: research and applications.
Yujie WU ; Jiarui XU ; Hongyu CHEN ; Hao DU
Chinese Journal of Biotechnology 2024;40(1):1-14
The fungal bioluminescence pathway (FBP) is a metabolic pathway responsible for the generation of bioluminescence derived from fungi. This pathway utilizes caffeic acid as the substrate, generating a high-energy intermediate, and the decomposition of which yields green fluorescence with a wavelength of approximately 520 nm. The FBP is evolutionally conserved in luminescent fungal groups. Unlike other bioluminescent systems, the FBP is particularly suitable for engineering applications in eukaryotic organisms, especially in plants. Currently, metabolically engineered luminescent plants are able to emit visible light to illuminate its surroundings, which can be visualized clearly in the dark. The fungal bioluminescent system could be explored in various applications in molecular biology, biosensors and glowing ornamental plants, and even green lighting along city streets.
Luminescence
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Light
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Fluorescence
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Eukaryota
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Green Light
4.Advances on BTB protein ubiquitination mediated plant development and stress response.
Tongtong LÜ ; Wenhui YAN ; Yan LIANG ; Yin DING ; Qingxia YAN ; Jinhua LI
Chinese Journal of Biotechnology 2024;40(1):63-80
The BTB (broad-complex, tramtrack, and bric-à-brac) domain is a highly conserved protein interaction motif in eukaryotes. They are widely involved in transcriptional regulation, protein degradation and other processes. Recently, an increasing number of studies have shown that these genes play important roles in plant growth and development, biotic and abiotic stress processes. Here, we summarize the advances of these proteins ubiquitination-mediated development and abiotic stress responses in plants based on the protein structure, which may facilitate the study of this type of gene in plants.
Eukaryota
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Plant Development/genetics*
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Proteolysis
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Ubiquitination
5.Evolutionary Signature of Information Transfer Complexity in Cellular Membrane Proteomes.
Jong Min KIM ; Byung Gee KIM ; S June OH
Genomics & Informatics 2009;7(2):111-121
Cell membrane proteins play crucial roles in the cell's molecular interaction with its environment and within itself. They consist of membrane-bound proteins and many types of transmembrane (TM) proteins such as receptors, transporters, channel proteins, and enzymes. Membrane proteomes of cellular organisms reveal some characteristics in their global topological distribution according to their evolutionary positions, and show their own information transfer complexity. Predicted transmembrane segments (TMSs) in membrane proteomes with HMMTOP showed near power-law distribution and frequency characteristics in 6-TMS and 7-TMS proteins in prokaryotes and eukaryotes, respectively. This reaffirms the important roles of membrane receptors in cellular communication and biological evolutionary history.
Eukaryota
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Membrane Proteins
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Membranes
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Proteins
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Proteome
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Signal Transduction
6.Inhibitory Effect of Eukaryotic Expression Vector Bearing TFPI-2 Gene on SHI-1 Cell Growth.
Jun-Jun LI ; Pei LIAO ; Feng WEN ; Ze-Yu LUO ; Yi-Xiong CAO
Journal of Experimental Hematology 2019;27(6):1812-1819
OBJECTIVE:
To construct a eukaryotic expression vector of human tissue factor pathway inhibitor-2 (TFPI-2) and to investigate the effect of TFPI-2 gene on the growth of acute monocytic leukemia cell line (SHI-1).
METHODS:
The cDNA of TFPI-2 was obtained by genetic chemical synthesis, the TFPI-2 gene and the linear vector fragment were ligated and inserted into the multiple cloning site of PEGFP-N1 vector, and the eukaryotic expression vector PEGFP-N1-TFPI-2 was transfected SHI-1 cells, then the obtained SHI-1 cells was observed by fluorescence microscopy; MTT assay was used to detect the effect of TFPI-2 gene on the relative growth rate of SHI-1 cells at the different time-point; RT-PCR was used to detect TFPI-2 mRNA expression levels in the cells of each group before and after TFPI-2 transfection; TFPI-2 protein expression was detected by Western blot. The cells which successfully transfected with PEGFP-N1-TFPI-2 vector were named as SHI-1-TFPI-2 (experimental group), and the cells transfected with the empty vector pEGFP-N1 and the untransfected cells were named as SHI-1-V and SHI-1-P and used as the control group.
RESULTS:
The human TFPI-2 gene eukaryotic expression vector PEGFP-N1-TFPI-2 was successfully constructed, then the transfected into SHI-1 cells, observed by fluorescence microscopy 24 hours later, as a result, the PEGFP-N1-TFPI-2 was successfully transferred into SHI-1 cells, and the number of fluorescent cells increased after 48 h and 72 h. RT-PCR showed that the gray scale ratio of TFPI-2 gene to β- actin in the experimental group was higher than that in the control group. The gray scale ratio was 0.51±0.04 in SHI-1-V group, 0.52±0.03 in SHI-1-P group, 0.87±0.08 in SHI-1-TFPI-2 group, and the difference between SHI-1-TFPI-2 and SHI-1-V, SHI-1-P group was statistically significant (P<0.05).
CONCLUSION
The expression of TFPI-2 gene in PEGFP-N1-TFPI-2 can inhibit the growth of SHI-1 cells, which provides a research direction for gene therapy of leukemia in the future.
Eukaryota
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Genetic Vectors
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Glycoproteins
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metabolism
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Green Fluorescent Proteins
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Humans
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Transfection
7.Group Ⅱ introns and the application in biotechnology: a review.
Guzhen CUI ; Dengxiong HUA ; Junying GU ; Zhenghong CHEN
Chinese Journal of Biotechnology 2022;38(3):915-924
Group Ⅱ introns are self-splicing ribozymes, which insert directly into target sites in DNA with high frequency through "retrohoming". They specifically and efficiently recognize and splice DNA target sites, endowing themselves with great potential in genetic engineering. This paper reviewed the gene targeting principle of group Ⅱ introns and the application in microbial genetic modification, and then analyzed the limitations of them in multi-functional gene editing and eukaryotes based on the "retrohoming" characteristics and the dependence on high Mg2+ concentration. Finally, we dissected the potential of group Ⅱ introns in the development of novel gene editing tools based on our previous research outcome and the structural characteristics of the introns, hoping to provide a reference for the application of group Ⅱ introns in biotechnology.
DNA
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Eukaryota
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Gene Targeting
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Introns/genetics*
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RNA, Catalytic/genetics*
8.Exploitation and utilization of rich lipids-microalgae, as new lipids feedstock for biodiesel production--a review.
Donghui SONG ; Lijun HOU ; Dingji SHI
Chinese Journal of Biotechnology 2008;24(3):341-348
As a renewable energy sources to replace conventional fossil fuels, biodiesel fuels have been becoming increasingly requirements to global fuels market. Biodiesel derived from oil crops cannot realistically satisfy even more fraction of the raw material existing costs and soil competitive demand for its growth. Microalgae appear to be the advantage of costs that is capable of higher photosynthetic efficiency, larger biomass, faster growth compared to those of oil crops. Lipid content of many microalgae is usually 80% of its dry weight. Genetic microalgae with high-oil productivity by genetic manipulations are capable of making microalgal biodiesel economically competitive with petrodiesel through large-scale production of genetic microalgal biomass. As demonstrated here, the use of biodiesel fuels in home and abroad are currently introduced, and the cost advantage of microalgae as the raw material is analyzed; And moreover, the progress of microalgal genetic engineering in regulation of lipid metabolism and the problems in the construct of genetic microalgae strains as well as approaches for making microalgal biodiesel appear to be an important source of renewable fuel that has the potential to completely displace fossil diesel are discussed in this review.
Bioelectric Energy Sources
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trends
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Biotechnology
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methods
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Eukaryota
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chemistry
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genetics
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metabolism
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Fatty Acids
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analysis
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Gasoline
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Lipids
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analysis
9.Mechanisms and assessment of water eutrophication.
Xiao-e YANG ; Xiang WU ; Hu-lin HAO ; Zhen-li HE
Journal of Zhejiang University. Science. B 2008;9(3):197-209
Water eutrophication has become a worldwide environmental problem in recent years, and understanding the mechanisms of water eutrophication will help for prevention and remediation of water eutrophication. In this paper, recent advances in current status and major mechanisms of water eutrophication, assessment and evaluation criteria, and the influencing factors were reviewed. Water eutrophication in lakes, reservoirs, estuaries and rivers is widespread all over the world and the severity is increasing, especially in the developing countries like China. The assessment of water eutrophication has been advanced from simple individual parameters like total phosphorus, total nitrogen, etc., to comprehensive indexes like total nutrient status index. The major influencing factors on water eutrophication include nutrient enrichment, hydrodynamics, environmental factors such as temperature, salinity, carbon dioxide, element balance, etc., and microbial and biodiversity. The occurrence of water eutrophication is actually a complex function of all the possible influencing factors. The mechanisms of algal blooming are not fully understood and need to be further investigated.
Animals
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Environment
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Eukaryota
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growth & development
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isolation & purification
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Eutrophication
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Humans
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Water
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analysis
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Water Microbiology
10.Addendum and emendation of "China Medicinal Animal Fauna".
Luqi HUANG ; Junde LI ; Shihuan TANG
China Journal of Chinese Materia Medica 2010;35(21):2927-2930
UNLABELLEDTo carry out the " China Medicinal Animal Fauna" addenda and revision, with effective assessment, protection, utilization of medicinal animal resources, to promote sustainable modem research and application for medicinal animals and medical materials from animals.
METHODKeep the original "China Medicinal Animal Fauna" characteristics and peculiarities, combined with nearly 30-year research progress of zoology and medicinal animals, and author's long-standing and rich experience.
RESULTDevelop the addenda's general framework, addenda and revision contents, revision methods and technical routes of the "China Medicinal Animal Fauna".
CONCLUSIONBased on the research of medicinal animal resource system, fully use of modern molecular biology and other emerging science and technology, rich the scientific connotation of medicinal material from animal, will promote the research and use of medical material from animal to a new level.
Animals ; China ; Eukaryota ; chemistry ; classification ; genetics ; Medicine, Chinese Traditional ; Reference Books, Medical