Efficient and Cost-Reduced Glucoamylase Fed-Batch Production with Alternative Carbon Sources

( Hong Zhen Luo ),( Han Liu ),( Zhenni He ),( Cong Zhou ),( Zhong Ping Shi ) 한국미생물 · 생명공학회 2015 Journal of microbiology and biotechnology Vol.25 No.2

Glucoamylase is an important industrial enzyme. Glucoamylase production by industrial Aspergillus niger strain featured with two major problems: (i) empirical substrate feeding methods deteriorating the fermentation performance; and (ii) the high raw materials cost limiting the economics of the glucoamylase product with delegated specification. In this study, we first proposed a novel three-stage varied-rate substrate feeding strategy for efficient glucoamylase production in a 5 L bioreactor using the standard feeding medium, by comparing the changing patterns of the important physiological parameters such as DO, OUR, RQ, etc., when using different substrate feeding strategies. With this strategy, the glucoamylase activity and productivity reached higher levels of 11,000 U/ml and 84.6 U/ml/h, respectively. The performance enhancement in this case was beneficial from the following results: DO and OUR could be controlled at the higher levels (30%, 43.83 mmol/l/h), while RQ was maintained at a stable/lower level of 0.60 simultaneously throughout the fed-batch phase. Based on this three-stage varied-rate substrate feeding strategy, we further evaluated the economics of using alternative carbon sources, attempting to reduce the raw materials cost. The results revealed that cornstarch hydrolysate could be considered as the best carbon source to replace the standard and expensive feeding medium. In this case, the production cost of the glucoamylase with delegated specification (5,000 U/ml) could be saved by more than 61% while the product quality be ensured simultaneously. The proposed strategy showed application potential in improving the economics of industrial glucoamylase production.

Vγ1+ γδT Cells Are Correlated With Increasing Expression of Eosinophil Cationic Protein and Metalloproteinase-7 in Chronic Rhinosinusitis With Nasal Polyps Inducing the Formation of Edema

Luo-ying Yang,Xia Li,Wen-ting Li,Jian-cong Huang,Zhi-yuan Wang,Zi-zhen Huang,Li-hong Chang,Ge-hua Zhang 대한천식알레르기학회 2017 Allergy, Asthma & Immunology Research Vol.9 No.2

Purpose: We have found that expression of γδT cells is increased in pathological mucosa of chronic rhinosinusitis with nasal polyps (CRSwNP) compared with normal nasal mucosa. This increase is correlated with the infiltration of eosinophils in CRSwNP. Here, we investigated the expression of γδT cells, inflammation and tissue remodeling factors as well as their probable relationships in different types of chronic rhinosinusitis (CRS) in China. Methods: A total of 76 surgical tissue samples that included 43 CRSwNP samples (15 eosinophilic and 28 non-eosinophilic), 17 CRS samples without nasal polyps (CRSsNP), and 16 controls were obtained. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the mRNA expression levels of Vγ1+ γδT cells, Vγ4+ γδT cells, eosinophil cationic protein (ECP), interleukin (IL)-8, transforming growth factor (TGF)-β2, metalloproteinase (MMP)-7, tissue inhibitor of metalloproteinase (TIMP)-4 and hypoxia-inducible factor (HIF)-1α. Enzyme linked immunosorbent assay (ELISA) was used to measure the protein level of ECP and MMP-7 in CRSwNP. The eosinophils were counted and the level of edema was analyzed with HE staining. Results: The mRNA expression levels of the Vγ1 subset, ECP and MMP-7 were significantly increased in CRSwNP with histological characteristics of eosinophilic infiltration and edema. The expression of the Vγ1 gene in CRSwNP correlated positively with the expression of both ECP and MMP-7. No significant decreases in the mRNA expression levels of TGF-β2, TIMP-4 or HIF-1α were observed in the CRSwNP samples. The expression levels of Vγ1 gene, ECP and MMP-7 were significantly increased in eosinophilic CRSwNP compared to non-eosinophilic CRSwNP. Conclusions: Our results suggest the associations between Vγ1+ γδT cells, ECP and MMP-7 in CRSwNP, indicating that Vγ1+ γδT cells can induce the eosinophilic inflammation, which has a further effect on the formation of edema.

IN SITU PREPARATION AND INHIBITORY ACTIVITY OF HYDROXYAPATITE/SILVER NANOCOMPOSITE

HONG-ZHEN XIE,JIAN-DONG WANG,CHONG-XIAO LUO,JIN-KU LIU 성균관대학교(자연과학캠퍼스) 성균나노과학기술원 2012 NANO Vol.7 No.6

The hydroxyapatite (HAP) assembled microsphere chains/silver (Ag) nanocomposites were prepared by a facile in situ preparation method under hydrothermal condition. The crystal structure of products was characterized by X-ray di®raction (XRD) and the morphologies of products were investigated by transmission electron microscopy (TEM). The nanocomposites own large surface area and can hydrogen-bond to other substances by the hydroxyl group on the surface. The nanocomposites have good structural stability under the electrostatic e®ect of the silver nanoparticles and strong adsorbability of the HAP microsphere chains. The two substances combine to form special nanocomposite spheres. Specially, the HAP/Ag nanocomposites have high inhibitory activity and can be applied in the environment and medicine ¯elds. The nanocomposite structures can save the consumption of Ag materials.

Separation Technology, Thermodynamics : Study of adsorption behavior using activated carbon for removal of colored impurities from 30% caprolactam solution produced by means of SNIA-toluene-technology

( Hong Tao Zhang ),( Ping Yu ),( Zhen Yuan ),( Yun Bai Luo ) 한국화학공학회 2006 Korean Journal of Chemical Engineering Vol.23 No.3

MicroRNA-576-3p Inhibits Proliferation in Bladder Cancer Cells by Targeting Cyclin D1

Liang, Zhen,Li, Shiqi,Xu, Xin,Xu, Xianglai,Wang, Xiao,Wu, Jian,Zhu, Yi,Hu, Zhenghui,Lin, Yiwei,Mao, Yeqing,Chen, Hong,Luo, Jindan,Liu, Ben,Zheng, Xiangyi,Xie, Liping Korean Society for Molecular and Cellular Biology 2015 Molecules and cells Vol.38 No.2

MicroRNAs (miRNAs) are small, endogenous RNAs that play important gene-regulatory roles by binding to the imperfectly complementary sequences at the 3'-UTR of mRNAs and directing their gene expression. Here, we first discovered that miR-576-3p was down-regulated in human bladder cancer cell lines compared with the non-malignant cell line. To better characterize the role of miR-576-3p in bladder cancer cells, we over-expressed or down-regulated miR-576-3p in bladder cancer cells by transfecting with chemically synthesized mimic or inhibitor. The overexpression of miR-576-3p remarkably inhibited cell proliferation via G1-phase arrest, and decreased both mRNA and protein levels of cyclin D1 which played a key role in G1/S phase transition. The knock-down of miR-576-3p significantly promoted the proliferation of bladder cancer cells by accelerating the progression of cell cycle and increased the expression of cyclin D1. Moreover, the dual-luciferase reporter assays indicated that miR-576-3p could directly target cyclin D1 through binding its 3'-UTR. All the results demonstrated that miR-576-3p might be a novel suppressor of bladder cancer cell proliferation through targeting cyclin D1.

Big Data Acquisition and Analysis Platform for Intermodal Transport

Kai Xu,Hong Zhen,Yan Li,Luo Yue 보안공학연구지원센터 2016 International Journal of Database Theory and Appli Vol.9 No.12

This paper aims for the transparency and visualization of the international intermodal cargo transportation throughout its whole process, achieving a comprehensive monitoring on the multiple transportation means such as by ocean, by air, by land or by rail. Based on Internet-of-Things-based distributed data acquisition technology and the cloud-computing-based big data analysis technology, this paper gives out a Multimodal Monitoring technology that can uniformly solve the comprehensive management of multiple transportation vehicles, which includes a service functionality model, a network hierarchy model and a technology system model. By building a Generic Target Monitoring System, it proves the multimodal monitoring resolution is able to effectively monitor the multiple transportation means and provide a fair good database platform of later analysis, distribution and optimization of those vehicles.

The Retention Behavior of Ginsenosides in HPLC and Its Application to Quality Assessment of Radix Ginseng

Hu, Ping,Luo, Guo-An,Wang, Qing,Zhao, Zhong-Zhen,Wang, Wan,Jiang, Zhi-Hong 대한약학회 2008 Archives of Pharmacal Research Vol.31 No.10

This study systematically investigated the retention behavior of seven neutral ginsenosides $Rg_1$, Re, Rf, $Rb_1$, $Rb_2$, Rc, Rd, and an acidic ginsenoside $R_0$, the major pharmacologically active components of Radix Ginseng with RP-HPLC. The effects of solvent, pH value, ionic strength of the mobile phase, and column temperature were investigated using an octadecylsiloxane-bonded silica gel column. Based on the ginsenosides' retention characteristics, the concentration of acetonitrile and the gradient of the mobile phase needed to maintain the baseline separation of the major neutral ginsenosides in Radix Ginseng were theoretically predicted. Furthermore, the ionic strength of mobile-phase necessary to achieve good resolution of the neutral ginsenosides and acidic ginsenosides was carefully investigated. According to the results of the quantitative analysis of ginsenosides in eight batches of ginseng samples from different sources, the developed HPLC technique may be a valuable tool for the quality assessment of Radix Ginseng.

MicroRNA-576-3p Inhibits Proliferation in Bladder Cancer Cells by Targeting Cyclin D1

Liping Xie,Zhen Liang,Shiqi Li,Xin Xu,Xianglai Xu,Xiao Wang,Jian Wu,Yi Zhu,Zhenghui Hu,Yiwei Lin,Yeqing Mao,Hong Chen,Jindan Luo,Ben Liu,Xiangyi Zheng 한국분자세포생물학회 2015 Molecules and cells Vol.38 No.2

MicroRNAs (miRNAs) are small, endogenous RNAs that play important gene-regulatory roles by binding to the imperfectly complementary sequences at the 3 -UTR of mRNAs and directing their gene expression. Here, we first discovered that miR-576-3p was down-regulated in human bladder cancer cell lines compared with the non-malignant cell line. To better characterize the role of miR-576-3p in bladder cancer cells, we over-expressed or down-regulated miR-576-3p in bladder cancer cells by transfecting with chemically synthesized mimic or inhibitor. The overexpression of miR-576-3p remarkably inhibited cell proliferation via G1-phase arrest, and decreased both mRNA and protein levels of cyclin D1 which played a key role in G1/S phase transition. The knock-down of miR-576-3p significantly promoted the proliferation of bladder cancer cells by accelerating the progression of cell cycle and increased the expression of cyclin D1. Moreover, the dual-luciferase reporter assays indicated that miR-576-3p could directly target cyclin D1 through binding its 3 -UTR. All the results demonstrated that miR-576-3p might be a novel suppressor of bladder cancer cell proliferation through targeting cyclin D1.

The Retention Behavior of Ginsenosides in HPLC and Its Application to Quality Assessment of Radix Ginseng

Ping Hu,Guo-An Luo,Zhong-Zhen Zhao,Wan Wang,Qing Wang,Zhi-Hong Jiang 대한약학회 2008 Archives of Pharmacal Research Vol.31 No.10

This study systematically investigated the retention behavior of seven neutral ginsenosides Rg₁, Re, Rf, Rb₁, Rb₂, Rc, Rd, and an acidic ginsenoside R0, the major pharmacologically active components of Radix Ginseng with RP-HPLC. The effects of solvent, pH value, ionic strength of the mobile phase, and column temperature were investigated using an octadecylsiloxane-bonded silica gel column. Based on the ginsenosides’ retention characteristics, the concentration of acetonitrile and the gradient of the mobile phase needed to maintain the baseline separation of the major neutral ginsenosides in Radix Ginseng were theoretically predicted. Furthermore, the ionic strength of mobile-phase necessary to achieve good resolution of the neutral ginsenosides and acidic ginsenosides was carefully investigated. According to the results of the quantitative analysis of ginsenosides in eight batches of ginseng samples from different sources, the developed HPLC technique may be a valuable tool for the quality assessment of Radix Ginseng.

Wnt/β-Catenin Promotes the Osteoblastic Potential of BMP9 Through Down-Regulating Cyp26b1 in Mesenchymal Stem Cells

Yao Xin-Tong,Li Pei-pei,Liu Jiang,Yang Yuan-Yuan,Luo Zhen-Ling,Jiang Hai-Tao,He Wen-Ge,Luo Hong-Hong,Deng Yi-Xuan,He Bai-Cheng 한국조직공학과 재생의학회 2023 조직공학과 재생의학 Vol.20 No.5

BACKGROUND: All-trans retinoic acid (ATRA) promotes the osteogenic differentiation induced by bone morphogenetic protein 9 (BMP9), but the intrinsic relationship between BMP9 and ATRA keeps unknown. Herein, we investigated the effect of Cyp26b1, a critical enzyme of ATRA degradation, on the BMP9-induced osteogenic differentiation in mesenchymal stem cells (MSCs), and unveiled possible mechanism through which BMP9 regulates the expression of Cyp26b1. METHODS: ATRA content was detected with ELISA and HPLC–MS/MS. PCR, Western blot, and histochemical staining were used to assay the osteogenic markers. Fetal limbs culture, cranial defect repair model, and micro–computed tomographic were used to evaluate the quality of bone formation. IP and ChIP assay were used to explore possible mechanism. RESULTS: We found that the protein level of Cyp26b1 was increased with age, whereas the ATRA content decreased. The osteogenic markers induced by BMP9 were increased by inhibiting or silencing Cyp26b1 but reduced by exogenous Cyp26b1. The BMP9-induced bone formation was enhanced by inhibiting Cyp26b1. The cranial defect repair was promoted by BMP9, which was strengthened by silencing Cyp26b1 and reduced by exogenous Cyp26b1. Mechanically, Cyp26b1 was reduced by BMP9, which was enhanced by activating Wnt/b-catenin, and reduced by inhibiting this pathway. b-catenin interacts with Smad1/5/9, and both were recruited at the promoter of Cyp26b1. CONCLUSIONS: Our findings suggested the BMP9-induced osteoblastic differentiation was mediated by activating retinoic acid signalling, viadown-regulating Cyp26b1. Meanwhile, Cyp26b1 may be a novel potential therapeutic target for the treatment of bone-related diseases or accelerating bone-tissue engineering.