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Expression of bkt and bch genes from Haematococcus pluvialis in transgenic Chlamydomonas

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Science China(Life Sciences) 2014年第10期57卷 1028-1033页

作者： ZHENG KaiJing WANG ChaoGang XIAO Ming CHEN Jun LI JianCheng HU ZhangLi Shenzhen Key Laboratory of Marine Bioresource & Eco-environmental Science Shenzhen Engineering Laboratory for Marine Algal Biotechnology College of Life Science Shenzhen University

β-carotene ketolase and β-carotene hydroxylase encoded by bkt and bch, respectively, are key enzymes required for astaxanthin biosynthesis in Haematococcu pluvialis 34-1n. Two expression vectors containing cDNA sequences of bkt and bch were constructed and co-transformed into cell-wall-deficient Chlamydomonas reinhardtii CC-849. Transgenic algae were screened on TAP agar plates containing 10 gg mL 1 Zeomycin. PCR-Southern analysis showed that bkt and bch were integrated into the genomes of C. reinhardtii. Transcripts of bkt and bch were further confirmed by RT-PCR-Southern analysis. Compared with the wild type, transgenic algae produced 29.04% and 30.27% more carotenoids and xanthophylls, respectively. Moreover, the transgenic algae could accumulate 34% more astaxanthin than wild type. These results indicate that foreign bkt and bch genes were successfully translated into β-carotene ketolase and β-carotene hydroxylase, which were responsible for catalyzing the biosynthesis of astaxanthin in transgenic algae.

关键词： Chlamydomonas β-carotene ketolase β-carotene hydroxylase carotenoids xanthophylls astaxanthin

Hepatoprotective activity of brown alga Padina boergesenii against CCl_4 induced oxidative damage in Wistar rats

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Asian Pacific Journal of Tropical Medicine 2010年第9期3卷 696-701页

作者： Rajamani Karthikeyan ST Somasundaram T Manivasagam T Balasubramanian P Anantharaman Centre of Advanced Study in Marine Biology Annamalai UniversityParangipettai-608 502Tamil NaduIndia Department of Bio-chemistry and Bio-technology Annamalai UniversityChidambaram-608 002Tamil NaduIndia

Objective:To investigate the protective effects of brown alga Padina boergesenii（***） against carbon tetrachloride（CCl4） induced oxidative damage and liver fibrosis in ***: To assess the hepatic damage liver weight,the activities of TBARS level,glutathione,SOD,CAT and GPx in circulation and ***:The group of rats induced with CCl4 alone（2 mL/kg body weight）,showed noticeable increase in the liver weight and TBARS *** by, the level of antioxidant enzymes glutathione peroxidase,Superoxide dismutase and catalase was also significantly（P4 induced liver *** level of antioxidant enzymes and lipid peroxidation products was found to be significandy（Pcarotenoids in the relevant activity,total carotenoid content of the extract was analysed and found to be 0.59 mg/g fresh weight（FW）.Further,the histopathogical studies provide a supportive evidence for this study to show the protective nature of ***:The protective role of brown alga *** extract has confirmed its potential activity through its antioxidant sparing actions against CCl4 induced free radical ***,the possible mechanism of hepatoprotection is rather speculative at this stage and investigations are underway to isolate and characterize the bioactive compounds from ***.

关键词： Hepatotoxicity Padina boergesenii Carbon tetrachloride Liver fibrosis carotenoids

Cytotoxic effect of Spirulina platensis extracts on human acute leukemia Kasumi-1 and chronic myelogenous leukemia K-562 cell lines

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Asian Pacific Journal of Tropical Biomedicine 2017年第1期7卷 14-19页

作者： Flor Yohana Flores Hernandez Sanghamitra Khandual Inocencia Guadalupe Ramírez López Center for Investigation and Assistance in Technology Design for State of JaliscoC.P.45019ZapopanJaliscoMexico

Objective: To evaluate the cytotoxic effects of Spirulina platensis extracts on acute leukemia Kasumi-1 and chronic leukemia K-562 cancer cell ***: Various concentrations of Spirulina platensis extracts(0.25–50.00 mg/m L)obtained with different solvents were used to treat cell lines for 72 h. For cytotoxic effect studies, cell viability test with trypan blue solution, MTT assay and microscopic cytomorphological assessment were ***: Spirulina extract obtained with 70% ethanol showed significant cytotoxicity in K562 and Kasumi-1 cell lines. With trypan blue solution, IC_(50) values were found to be4.64 mg/m L for K-562 and 3.68 mg/m L for Kusumi-1 cell lines. Spirulina aqueous extract also showed cytotoxicity with trypan blue method, at a slightly higher dose; where IC_(50) values were 12.68 mg/m L for K-562 and 2.13 mg/m L for Kusumi-1 cell lines. The IC_(50) values were found 0.40 mg/m L for K-562 and 0.31 mg/m L for Kusumi-1 cell lines for the 70% ethanol extract according to the MTT assay. Spirulina extract obtained with water also showed cytotoxicity but the dose was a little higher where IC_(50) values were15.77 mg/m L for K-562 and 9.44 mg/m L for Kusumi-1 cell lines. The effect of cytotoxicity with ethanol extract is quite comparable with that observed for cyclophosphamide, which is a chemical used as anticancer ***: The cytotoxicity exhibited by Spirulina extract to cancer cell lines might be due to the presence of phytopigments(carotenoids, chlorophyll, phycocyanin) as well as polysaccharides that were reported previously as constituents of the extract. So crude extracts of Spirulina can be used as a source to develop anticancer drugs.

关键词： Leukemia Cytotoxicity Spirulina carotenoids

Optimization of microbial cell factories for astaxanthin production: Biosynthesis and regulations, engineering strategies and fermentation optimization strategies

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Synthetic and Systems Biotechnology 2022年第2期7卷 689-704页

作者： Mostafa Basiony Liming Ouyang Danni Wang Jiaming Yu Liming Zhou Mohan Zhu Xuyuan Wang Jie Feng Jing Dai Yijie Shen Chengguo Zhang Qiang Hua Xiuliang Yang Lixin Zhang State Key Laboratory of Bioreactor Engineering East China University of Science and TechnologyShanghai200237China Shandong Jincheng Bio-Pharmaceutical Co. Ltd.No.117 Qixing River RoadZibo255130ShandongChina

The global market demand for natural astaxanthin is rapidly increasing owing to its safety,the potential health benefits,and the diverse applications in food and pharmaceutical *** major native producers of natural astaxanthin on industrial scale are the alga Haematococcus pluvialis and the yeast Xanthopyllomyces ***,the natural production via these native producers is facing challenges of limited yield and high cost of cultivation and ***,astaxanthin production via metabolically engineered non-native microbial cell factories such as Escherichia coli,Saccharomyces cerevisiae and Yarrowia lipolytica is another promising strategy to overcome these *** this review we summarize the recent scientific and biotechnological progresses on astaxanthin biosynthetic pathways,transcriptional regulations,the interrelation with lipid metabolism,engineering strategies as well as fermentation process control in major native and non-native astaxanthin *** progresses illuminate the prospects of producing astaxanthin by microbial cell factories on industrial scale.

关键词： Astaxanthin carotenoids Metabolic engineering Microbial cell factories Lipid Fermentation Antioxidants

Molecular Cloning and Expression Analysis of the ζ-Carotene Desaturase Gene in Chinese kale(Brassica oleracea *** Bailey)

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Horticultural Plant Journal 2018年第3期4卷 94-102页

作者： SUN Bo ZHANG Fen XUE Shengling CHANG Jiaqi ZHENG Aihong JIANG Min MIAO Huiying WANG Qiaomei TANG Haoru College of Horticulture Sichuan Agricultural University Department of Horticulture Zhejiang University

ζ-Carotene desaturase(ZDS)is an important enzyme in carotenoid ***,the Brassica oleracea *** ZDS(Boa ZDS)gene was cloned from Chinese kale via reverse transcription-polymerase chain reaction(RT-PCR)and deposited in Gen Bank(accession number KY662297).The Boa ZDS gene contains an open reading frame of 1 686 bp that encodes a 561-amino acid *** analysis indicates that the ZDS protein is apparently conserved during plant evolution and is most closely related to *** *** and *** promoter sequence of the Boa ZDS gene was predicted to harbor several cis-acting elements that are related to light and phytohormone *** RT-PCR analysis showed that Boa ZDS expression varied among different developmental stages and *** ZDS expression remained stable during germination and seedling stages and rapidly increased at the mature leaf *** leaves showed the highest ZDS expression levels compared to the other *** expression decreased in all flower tissues during *** fused protein of Boa ZDS was obtained by prokaryotic *** expression of Boa ZDS in Escherichia coli confirmed that Boa ZDS encodes a functionalζ-carotene desaturase that increases β-carotene accumulation in *** cells harboring a β-carotene-producing *** findings of the present study provide a molecular basis for the elucidation of ZDS gene function in Chinese kale.

关键词： Chinese kale Brassica oleracea var alboglabra Bailey carotenoids ζ-carotene desaturase(ZDS) cloning gene expression prokaryotic expression heterologous expression

Ultrasound‑assisted osmotic dehydration,followed by convective drying of papaya:effect on physicochemical and functional quality parameters

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Systems Microbiology and Biomanufacturing 2023年第4期3卷 615-626页

作者： Abhishek Chandra Sourabh Kumar Sachin Kumar Prabhat Kumar Nema Department of Food Engineering National Institute of Food Technology Entrepreneurship and ManagementSonipat 131028India School of Health Sciences and Technology University of Petroleum and Energy StudiesDehradunUttarakhandIndia

Papaya is a rich source of nutrients,antioxidants,and vitamins;however,it is highly perishable and hence needs to be preserved using suitable unit *** is an economical method of papaya treatment,but results in the loss of essential nutrients and also degrades its pigments,color,and ***,this work aims to utilize ultrasound and osmotic dehydration as pretreatment strategies to preserve the rich composition of papaya during conventional hot air *** dried papaya slices were evaluated for moisture content,water activity,total phenolic content,β-carotene,color,antioxidant activity,microstructure,and *** showed that sample pretreatment with 33 kHz ultrasound frequency and dehydrated with 35°Brix osmotic solution resulted in lower moisture content(<12%w.b.)and water activity(<0.41),high phenolic content(88.5 mg GAE/mL),highβ-carotene content(184.54 mg/g)and antioxidant capacity(48.3%),and preserved color after *** texture(hardness)of the pretreated samples was also more desirable from an industrial standpoint,which was further validated using SEM micrographs that showed a more porous *** study concludes that pretreatment of papaya slices with ultrasound and osmotic dehydration is recommended before drying.

关键词： Carica papaya Microstructure carotenoids Texture Drying Pretreatment

Pressurized Liquid Extraction of Pigments from Chlamydomonas *** Chemical Characterization by HPLC–MS/MS

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Journal of Analysis and Testing 2018年第2期2卷 149-157页

作者： Lidia Montero Mahshid Sedghi Yolanda García Carlos Almeida Carl Safi Nicole Engelen‑Smit Alejandro Cifuentes Jose A.Mendiola Elena Ibáñez Applied Analytical Chemistry University of Duisburg-EssenUniversitaetsstr.545141 EssenGermany Laboratory of Foodomics Bioactivity and Food Analysis DepartmentInstitute of Food Science Research CIAL(CSIC-UAM)Campus de CantoblancoC/Nicolás Cabrera 928049 MadridSpain Spanish Bank of Algae Instituto de Oceanografía y Cambio GlobalFundación Canaria Parque Científico‑TecnológicoUniversidad de Las Palmas de Gran CanariaMuelle de Taliarte s/n35214 Telde(Canary Islands)Spain Wageningen Food and Biobased Research Wageningen University and ResearchPO Box 176700 AA WageningenThe Netherlands

Pressurized liquid extraction using green solvents(ethanol and water)was applied to extract photosynthetic pigments from the microalgae Chlamydomonas *** experimental design is employed to determine the effect of the extraction conditions on the extraction yield and the antioxidant capacity of the different extracts determined using 1,1-diphenyl-2-picryl-hydrazyl and TEAC *** the extracts show significant antioxidant activity in all ***,in this work,the carotenoid and chlorophyll composition of the Chlamydomonas extracts was analyzed by HPLC–DAD–APCI–MS/MS for the first time,being the main compounds lutein and pheophythin a′.

关键词： Antioxidant capacity Chlamydomonas sp. HPLC–DAD–APCI–MS/MS Pigments Pressurized liquid extraction carotenoids

Cloning, Expression of Crocus sativus Phytoene Desaturase Gene and Preparation of Antiserum against It

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Wuhan University Journal of Natural Sciences 2004年第2期9卷 252-258页

作者： BaiJie MiaoChen XuYing TangLin WangZhi-tao ChenFang CollegeofLifeSciences SichuanUniversityChengdu610064SiehuanChina

A 2 149 bp full length phytoene desaturase (PDS) cDNA was first cloned from saffron (Crocus sativus L.) stigma using RT-PCR technique and a rapid amplification of cDNA end (RACE) strategy. The cDNA has an open reading frame of 1 697 bp, which encodes a polypeptide of 565 amino acids. The coding region of the cDNA was inserted into a prokaryotic expression vector pET-21a(+) and over-expressed inE. coli BL21 (DE3). The fusion proteins were found largely in an insoluble inclusion bodies. The purified fusion protein was used to immunize rabbits to obtain polyclonal antiserum with titer of 1×105. Western blot analysis by using this particular antiserum showed that the higher expression level of PDS in mature stigma than in leaves and stamen, and the higher expression level of PDS in mature stigma than in young stigma. Key words saffron - carotenoids - phytoene desaturase - gene expression - antiserum - Western blot CLC number Q 781 - Q 786 Foundation item: Supported by the Doctoral Foundation of the Ministry of Education, P. R. China and the Young Science Foundation of Sichuan University (Grant 0020405505012)Biography: Bai Jie (1968-), female, Ph. D candidate, research direction: plant developmental biology and reproductive engineering.

关键词： saffron carotenoids phytoene desaturase gene expression antiserum Western blot

Corrigendum:Edible seaweed-derived constituents:an undisclosed source of neuroprotective compounds

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Neural Regeneration Research 2021年第12期16卷 2564-2568页

作者： Melissa Schepers Nikita Martens Assia Tiane Kenneth Vanbrabant Hong-Bing Liu Dieter Lütjohann Monique Mulder Tim Vanmierlo Department of Neuroimmunology Biomedical Research InstituteHasselt UniversityEuropean Graduate School of Neuroscience(EURON)HasseltBelgium Department of Psychiatry&Neuropsychology Division of Translational NeuroscienceSchool for Mental Health and NeuroscienceMaastricht UniversityEuropean Graduate School of Neuroscience(EURON)MaastrichtThe Netherlands Department of Internal Medicine Laboratory of Vascular MedicineErasmus MC University Medical CenterRotterdamthe Netherlands Institue for Clinical Chemistry and Clinical Pharmacology BonnGermany Key Laboratory of Marine Drugs Ministry of EducationSchool of Medicine and PharmacyOcean University of ChinaQingdaoShandong ProvinceChina

Edible marine algae,or seaweeds,are a rich source of several bioactive compounds including phytosterols,carotenoids,and *** the last decades,seaweedderived constituents turned out to not only reside in the systemic circulation,but are able to cross the blood-brain barrier to exert neuro-active functions both in homeostatic and pathological ***,seaweed-derived constituents have gained increasing interest for their neuro-immunomodulatory and neuroprotective properties,rendering them interesting candidates for the management of several neurodegenerative *** particular seaweed-derived phytosterols gained interest for the treatment of neurodegenerative disorders as they potentiate neuroplasticity,enhance phagocytic clearance of neurotoxic peptides and have anti-inflammatory ***,the antiinflammatory and anti-oxidative properties of other constituents including carotenoids,phenols and polysaccharides have recently gained more *** this review,we provide an overview of a selection of the described neuro-active properties of seaweed-derived constituents with a focus on phytosterols.

关键词： algae carotenoids neuro-active phenols phytosterols polysaccharides seaweed