<i>Roseivivax</i> <i>roseus</i> sp. nov., an alphaproteobacterium isolated from a solar saltern soil sample

Zhang, Yu-Qin,Lee, Jae-Chan,Park, Dong-Jin,Lu, Xin-Xin,Mou, Xiao-Zhen,Kim, Chang-Jin International Union of Microbiological Societies 2014 International journal of systematic and evolutiona Vol.64 No.5

<P>A pink, Gram-stain-negative, motile, halotolerant bacterium with subpolar flagellum, designated strain BH87090<SUP>T</SUP>, was isolated from a saline soil sample collected from the south-west coastal area of South Korea (125° 58′ 58.08″ E 34° 45′ 37.32″ N). The isolate formed opaque pink to red colonies on marine agar plates at 30 °C. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, sulfoquinovosyl diacylglycerol, phosphatidylcholine and one unidentified phospholipid. The sole respiratory quinone was ubiquinone-10 (Q-10). The major cellular fatty acids were C<SUB>18 : 1</SUB>ω7<I>c</I>, C<SUB>19 : 0</SUB> cyclo ω8<I>c</I>, C<SUB>16 : 0</SUB> and 11-methyl C<SUB>18 : 1</SUB>ω7<I>c</I>. The genomic DNA G+C content was 61.8 mol%. These chemotaxonomic characteristics were all consistent with specific properties of the genus <I>Roseivivax</I>. Phylogenetic analysis based on 16S rRNA gene sequences showed that the isolate affiliated to the cluster with members of the genus <I>Roseivivax</I> in the <I>Roseobacter</I> clade, which suggested that the strain belonged to the genus <I>Roseivivax</I>. However, the low 16S rRNA gene similarities (93.5–95.3 %) of strain BH87090<SUP>T</SUP> with all the members of the genus <I>Roseivivax</I> indicated that it represented a novel species of the genus <I>Roseivivax</I>. On the basis of phenotypic and genotypic data, strain BH87090<SUP>T</SUP> should be classified as a novel species of the genus <I>Roseivivax</I>. The name <I>Roseivivax roseus</I> sp. nov. is proposed, with strain BH87090<SUP>T</SUP> ( = DSM 23042<SUP>T</SUP> = KCTC 22650<SUP>T</SUP>) as the type strain.</P>

A Three-step Proteolytic Cascade Mediates the Activation of the Peptidoglycan-induced Toll Pathway in an Insect

Kim, Chan-Hee,Kim, Su-Jin,Kan, Hongnan,Kwon, Hyun-Mi,Roh, Kyung-Baeg,Jiang, Rui,Yang, Yu,Park, Ji-Won,Lee, Hyeon-Hwa,Ha, Nam-Chul,Kang, Hee Jung,Nonaka, Masaru,Sö,derhä,ll, Kenneth,Lee, Bok Lu American Society for Biochemistry and Molecular Bi 2008 The Journal of biological chemistry Vol.283 No.12

<P>The recognition of lysine-type peptidoglycans (PG) by the PG recognition complex has been suggested to cause activation of the serine protease cascade leading to the processing of Spätzle and subsequent activation of the Toll signaling pathway. So far, two serine proteases involved in the lysine-type PG Toll signaling pathway have been identified. One is a modular serine protease functioning as an initial enzyme to be recruited into the lysine-type PG recognition complex. The other is the Drosophila Spätzle processing enzyme (SPE), a terminal enzyme that converts Spätzle pro-protein to its processed form capable of binding to the Toll receptor. However, it remains unclear how the initial PG recognition signal is transferred to Spätzle resulting in Toll pathway activation. Also, the biochemical characteristics and mechanism of action of a serine protease linking the modular serine protease and SPE have not been investigated. Here, we purified and cloned a novel upstream serine protease of SPE that we named SAE, SPE-activating enzyme, from the hemolymph of a large beetle, Tenebrio molitor larvae. This enzyme was activated by Tenebrio modular serine protease and in turn activated the Tenebrio SPE. The biochemical ordered functions of these three serine proteases were determined in vitro, suggesting that the activation of a three-step proteolytic cascade is necessary and sufficient for lysine-type PG recognition signaling. The processed Spätzle by this cascade induced antibacterial activity in vivo. These results demonstrate that the three-step proteolytic cascade linking the PG recognition complex and Spätzle processing is essential for the PG-dependent Toll signaling pathway.</P>

Engineering Bifunctional Antibodies with Constant Region Fusion Architectures

Du, Juanjuan,Cao, Yu,Liu, Yan,Wang, Ying,Zhang, Yong,Fu, Guangsen,Zhang, Yuhan,Lu, Lucy,Luo, Xiaozhou,Kim, Chan Hyuk,Schultz, Peter G.,Wang, Feng American Chemical Society 2017 JOURNAL OF THE AMERICAN CHEMICAL SOCIETY - Vol.139 No.51

<P>We report a method to generate bifunctional antibodies by grafting full-length proteins into constant region loops of a full-length antibody or an antigen-binding fragment (Fab). The fusion proteins retain the antigen binding activity of the parent antibody but have an additional activity associated with the protein insert. The engineered antibodies have excellent <I>in vitro</I> activity, physiochemical properties, and stability. Among these, a Her2 × CD3 bispecific antibody (BsAb) was constructed by inserting an anti-Her2 single-chain variable fragment (ScFv) into an anti-CD3 Fab. This bispecific antibody efficiently induces targeted cell lysis in the presence of effector cells at as low as sub-picomolar concentrations <I>in vitro</I>. Moreover, the Her2 × CD3 BsAb shows potent <I>in vivo</I> antitumor activity in mouse Her2<SUP>2+</SUP> and Her2<SUP>1+</SUP> xenograft models. These results demonstrate that insertion of a full-length protein into non-CDR loops of antibodies provides a feasible approach to generate multifunctional antibodies for therapeutic applications.</P><P><B>Graphic Abstract</B> <IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/2017/jacsat.2017.139.issue-51/jacs.7b09641/production/images/medium/ja-2017-09641x_0004.gif'></P><P><A href='http://pubs.acs.org/doi/suppl/10.1021/ja7b09641'>ACS Electronic Supporting Info</A></P>

Minimum Risk Capital in Project Finance: Example of Harbor Infrastructure Development

( Chen Kang Shu ),( Chan Yu Lu ) 한국해운물류학회(구 한국해운학회) 2007 해운물류연구 Vol.54 No.-

Assessing Neurobehavioral Alterations Among E-waste Recycling Workers in Hong Kong

Liao Gengze,Wang Feng,Lu Shaoyou,Yu Yanny Hoi Kuen,Arrandale Victoria H.,Chan Alan Hoi-shou,Tse Lap Ah 한국산업안전보건공단 산업안전보건연구원 2024 Safety and health at work Vol.15 No.1

Background E-waste workers in Hong Kong are handling an unprecedented amount of e-waste, which contains various neurotoxic chemicals. However, no study has been conducted to evaluate the neurological health status of e-waste workers in Hong Kong. This study aimed to evaluate the prevalence of neurobehavioral alterations and to identify the vulnerable groups among Hong Kong e-waste workers. Methods We recruited 109 Hong Kong e-waste workers from June 2021 to September 2022. Participants completed standard questionnaires and wore a GENEActiv accelerometer for seven days. Pittsburgh Sleep Quality Index and Questionnaire 16/18 (Q16/18) were used to assess subjective neurobehavioral alterations. The GENEActiv data generated objective sleep and circadian rhythm variables. Workers were grouped based on job designation and entity type according to the presumed hazardous level. Unconditional logistic regression models measured the associations of occupational characteristics with neurobehavioral alterations after adjusting for confounders. Results While dismantlers/repairers and the workers in entities not funded by the government were more likely to suffer from neurotoxic symptoms in Q18 (adjusted odds ratio: 3.18 [1.18–9.39] and 2.77 [1.10–7.46], respectively), the workers from self-sustained recycling facilities also have poor performances in circadian rhythm. Results also showed that the dismantlers/repairers working in entities not funded by the government had the highest risk of neurotoxic symptoms compared to the lowest-risk group (i.e., workers in government-funded companies with other job designations). Conclusion This timely and valuable study emphasizes the importance of improving the working conditions for high-risk e-waste workers, especially the dismantlers or repairers working in facilities not funded by the government. Background E-waste workers in Hong Kong are handling an unprecedented amount of e-waste, which contains various neurotoxic chemicals. However, no study has been conducted to evaluate the neurological health status of e-waste workers in Hong Kong. This study aimed to evaluate the prevalence of neurobehavioral alterations and to identify the vulnerable groups among Hong Kong e-waste workers. Methods We recruited 109 Hong Kong e-waste workers from June 2021 to September 2022. Participants completed standard questionnaires and wore a GENEActiv accelerometer for seven days. Pittsburgh Sleep Quality Index and Questionnaire 16/18 (Q16/18) were used to assess subjective neurobehavioral alterations. The GENEActiv data generated objective sleep and circadian rhythm variables. Workers were grouped based on job designation and entity type according to the presumed hazardous level. Unconditional logistic regression models measured the associations of occupational characteristics with neurobehavioral alterations after adjusting for confounders. Results While dismantlers/repairers and the workers in entities not funded by the government were more likely to suffer from neurotoxic symptoms in Q18 (adjusted odds ratio: 3.18 [1.18–9.39] and 2.77 [1.10–7.46], respectively), the workers from self-sustained recycling facilities also have poor performances in circadian rhythm. Results also showed that the dismantlers/repairers working in entities not funded by the government had the highest risk of neurotoxic symptoms compared to the lowest-risk group (i.e., workers in government-funded companies with other job designations). Conclusion This timely and valuable study emphasizes the importance of improving the working conditions for high-risk e-waste workers, especially the dismantlers or repairers working in facilities not funded by the government.

Engineering a short, aldolase-based pathway for (<i>R</i>)-1,3-butanediol production in <i>Escherichia coli</i>

Nemr, Kayla,Mü,ller, Jonas E.N.,Joo, Jeong Chan,Gawand, Pratish,Choudhary, Ruhi,Mendonca, Burton,Lu, Shuyi,Yu, Xiuyan,Yakunin, Alexander F.,Mahadevan, Radhakrishnan Elsevier 2018 Metabolic engineering Vol.48 No.-

<P><B>Abstract</B></P> <P>Microbial processes can produce a wide range of compounds; however, producing complex and long chain hydrocarbons remains a challenge. Aldol condensation offers a direct route to synthesize these challenging chemistries and can be catalyzed by microbes using aldolases. Deoxyribose-5-phosphate aldolase (DERA) condenses aldehydes and/or ketones to β -hydroxyaldehydes, which can be further converted to value-added chemicals such as a precursor to cholesterol-lowering drugs. Here, we implement a short, aldolase-based pathway in <I>Escherichia coli</I> to produce (<I>R</I>)-1,3-BDO from glucose, an essential component of pharmaceutical products and cosmetics. First, we expressed a three step heterologous pathway from pyruvate to produce 0.3 g/L of (<I>R</I>)-1,3-BDO with a yield of 11.2 mg/g of glucose in wild-type <I>E. coli</I> K12 MG1655. We used a systems metabolic engineering approach to improve (<I>R</I>)-1,3-BDO titer and yield by: 1) identifying and reducing major by-products: ethanol, acetoin, and 2,3-butanediol; 2) increasing pathway flux through DERA to reduce accumulation of toxic acetaldehyde. We then implemented a two-stage fermentation process to improve (<I>R</I>)-1,3-BDO titer by 8-fold to 2.4 g/L and yield by 5-fold to 56 mg/g of glucose ( 11 % of maximum theoretical yield) in strain BD24, by controlling pH to 7 and higher dissolved oxygen level. Furthermore, this study highlights the potential of the aldolase chemistry to synthesize diverse products directly from renewable resources in microbes.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Platform for non-natural chemicals developed using aldol condensation. </LI> <LI> Modular pathway design demonstrated in E. coli for (R)-1,3-BDO production. </LI> <LI> Carbon flux optimized by blocking pyruvate and acetaldehyde-consuming pathways. </LI> <LI> Final (R)-1,3-BDO production: 2.4 g/L and 11% of maximum theoretical yield. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>