Cloning, Heterologous Expression, and Characterization of Novel Protease- Resistant α-Galactosidase from New Sphingomonas Strain

( Jun Pei Zhou ),( Yan Yan Dong ),( Jun Jun Li ),( Rui Zhang ),( Xianghua Tang ),( Yuelin Mu ),( Bo Xu ),( Qian Wu ),( Zun Xi Huang ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.11

The α-galactosidase-coding gene agaAJB13 was cloned from Sphingomonas sp. JB13 showing 16S rDNA (1,343 bp) identities of ≤97.2% with other identified Sphingomonas strains. agaAJB13 (2,217 bp; 64.9% GC content) encodes a 738-residue polypeptide (AgaAJB13) with a calculated mass of 82.3 kDa. AgaAJB13 showed the highest identity of 61.4% with the putative glycosyl hydrolase family 36 α-galactosidase from Granulicella mallensis MP5ACTX8 (EFI56085). AgaAJB13 also showed <37% identities with reported protease-resistant or Sphingomonas α-galactosidases. A sequence analysis revealed different catalytic motifs between reported Sphingomonas α-galactosidases (KXD and RXXXD) and AgaAJB13 (KWD and SDXXDXXXR). Recombinant AgaAJB13 (rAgaAJB13) was expressed in Escherichia coli BL21 (DE3). The purified rAgaAJB13 was characterized using p-nitrophenyl-α-D-galactopyranoside as the substrate and showed an apparent optimum at pH 5.0 and 60oC and strong resistance to trypsin and proteinase K digestion. Compared with reported proteaseresistant α-galactosidases showing thermolability at 50oC or 60oC and specific activities of <71 U/mg with or without protease treatments, rAgaAJB13 exhibited a better thermal stability (half-life of >60 min at 60oC) and higher specific activities (225.0-256.5 U/mg). These sequence and enzymatic properties suggest AgaAJB13 is the first identified and characterized Sphingomonas α-galactosidase, and shows novel protease resistance with a potential value for basic research and industrial applications.

Production and Characterization of Ethanol- and Protease-Tolerant and Xylooligosaccharides-Producing Endoxylanase from Humicola sp Ly01

( Jun Pei Zhou ),( Qian Wu ),( Rui Zhang ),( Yu Ying Yang ),( Xiang Hua Tang ),( Jun Jun Li ),( Jun Mei Ding ),( Yan Yan Dong ),( Zun Xi Huang ) 한국미생물 · 생명공학회 2013 Journal of microbiology and biotechnology Vol.23 No.6

This paper reports the production and characterization of crude xylanase from the newly isolated Humicola sp. Ly01. The highest (41.8 U/ml) production of the crude xylanase was obtained under the optimized conditions (w/v): 0.5% wheat bran, 0.2% KH2PO4, and 0.5% peptone; initial pH 7.0; incubation time 72 h; 30℃; and 150 rpm. A considerable amount of the crude xylanase was induced using hulless barley bran or soybean meal as the carbon source, but a small amount of the enzyme was produced when supplementary urea was used as the nitrogen source to wheat bran. The crude xylanase showed apparent optimal cellulase-free xylanase activity at 60℃ and pH 6.0, more than 71.8% of the maximum xylanase activity in 3.0-30.0% (v/v) ethanol and more than 82.3% of the initial xylanase activity after incubation in 3.0-30.0% (v/v) ethanol at 30℃ for 2 h. The crude xylanase was moderately resistant to both acid and neutral protease digestion, and released 7.9 and 10.9 μmol/ml reducing sugar from xylan in the simulated gastric and intestinal fluids, respectively. The xylooligosaccharides were the main products of the hydrolysis of xylan by the crude xylanase. These properties suggested the potential of the crude enzyme for being applied in the animal feed industry, xylooligosaccharides production, and high-alcohol conditions such as ethanol production and brewing.

Molecular and Biochemical Characterization of a Novel Intracellular Low-Temperature-Active Xylanase

( Jun Pei Zhou,),( Yan Yan Dong ),( Xiang Hua Tang ),( Jun Jun Li ),( Bo Xu ),( Qian Wu ),( Ya Jie Gao ),( Lu Pan ),( Zun Xi Huang ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.4

A 990 bp full-length gene (xynAHJ2) encoding a 329- residue polypeptide (XynAHJ2) with a calculated mass of 38.4 kDa was cloned from Bacillus sp. HJ2 harbored in a saline soil. XynAHJ2 showed no signal peptide, distinct amino acid stretches of glycoside hydrolase (GH) family 10 intracellular endoxylanases, and the highest amino acid sequence identity of 65.3% with the identified GH 10 intracellular mesophilic endoxylanase iM-KRICT PX1-Ps from Paenibacillus sp. HPL-001 (ACJ06666). The recombinant enzyme (rXynAHJ2) was expressed in Escherichia coli and displayed the typical characteristics of low-temperatureactive enzyme (exhibiting optimum activity at 35 o C, 62% at 20 o C, and 38% at 10 o C; thermolability at ≥45 o C). Compared with the reported GH 10 low-temperature-active endoxylanases, which are all extracellular, rXynAHJ2 showed low amino acid sequence identities (<45%), low homology (different phylogenetic cluster), and difference of structure (decreased amount of total accessible surface area and exposed nonpolar accessible surface area). Compared with the reported GH 10 intracellular endoxylanases, which are all mesophilic and thermophilic, rXynAHJ2 has decreased numbers of arginine residues and salt bridges, and showed resistance to Ni 2+ , Ca 2+ , or EDTA at 10 mM final concentration. The above mechanism of structural adaptation for low-temperature activity of intracellular endoxylanase rXynAHJ2 is different from that of GH 10 extracellular low-temperature-active endoxylanases. This is the first report of the molecular and biochemical characterizations of a novel intracellular low-temperatureactive xylanase.

Biochemical Characterization of a GDSL-Motif Esterase from Bacillus sp. K91 with a New Putative Catalytic Mechanism

( Jun Mei Ding ),( Ting Ting Yu ),( Lian Ming Liang ),( Zhen Rong Xie ),( Yun Juan Yang ),( Jun Pei Zhou ),( Bo Xu ),( Jun Jun Li ),( Zun Xi Huang ) 한국미생물 · 생명공학회 2014 Journal of microbiology and biotechnology Vol.24 No.11

The esterase gene Est8 from the thermophilic bacterium Bacillus sp. K91 was cloned and expressed in Escherichia coli. The monomeric enzyme exhibited a theoretical molecular mass of 24.5 kDa and an optimal activity around 50°C at pH 9.0. A model of Est8 was constructed using a hypothetical YxiM precursor structure (2O14_A) from Bacillus subtilis as template. The structure showed an α/β-hydrolase fold and indicated the presence of a typical catalytic triad consisting of Ser-11, Asp-182, and His-185, which were investigated by site directed replacements coupled with kinetic characterization. Asp-182 and His-185 residues were more critical than the Ser-11 residue in the catalytic activity of Est8. A comparison of the amino acid sequence showed that Est8 could be grouped into the GDSL family and further classified as an SGNH hydrolase. Est8 is a new member of the SGNH hydrolase subfamily and may employ a different catalytic mechanism.

Zoster sine herpete: a review

( Junli Zhou ),( Juan Li ),( Lulin Ma ),( Song Cao ) 대한통증학회 2020 The Korean Journal of Pain Vol.33 No.3

Zoster sine herpete (ZSH) is one of the atypical clinical manifestations of herpes zoster (HZ), which stems from infection and reactivation of the varicella-zoster virus (VZV) in the cranial nerve, spinal nerve, viscera, or autonomic nerve. Patients with ZSH display variable symptoms, such as neuralgia, however, different from HZ, ZSH show no zoster, which makes clinical diagnosis difficult. ZSH not only causes initial symptoms, such as neuropathic pain in the affected nerve, Bell palsy, and Ramsay Hunt syndrome, but also postherpetic neuralgia and fatal complications such as VZV encephalitis and stroke. The misdiagnosis of ZSH and tardy antiviral treatment may lead to severe ZSH sequelae. We review the publications related to ZSH, especially its diagnosis with VZV DNA and/or anti-VZV immunoglobulin (IgG and IgM). More work about ZSH, especially ZSH epidemiological survey and guidelines for its diagnosis and treatment, are needed because most of the present studies are case reports.

BIFURCATION ANALYSIS OF A SINGLE SPECIES REACTION-DIFFUSION MODEL WITH NONLOCAL DELAY

Zhou, Jun Korean Mathematical Society 2020 대한수학회지 Vol.57 No.1

A reaction-diffusion model with spatiotemporal delay modeling the dynamical behavior of a single species is investigated. The parameter regions for the local stability, global stability and instability of the unique positive constant steady state solution are derived. The conditions of the occurrence of Turing (diffusion-driven) instability are obtained. The existence of time-periodic solutions, the existence and nonexistence of nonconstant positive steady state solutions are proved by bifurcation method and energy method. Numerical simulations are presented to verify and illustrate the theoretical results.

Development of action research in China: review and reflection

Jun Zhou,Katrina Yan Liua 서울대학교 교육연구소 2011 Asia Pacific Education Review Vol.12 No.2

Action research in China during late 1970s and 1980s was influenced by positivist scientific research, believing the purpose of doing action research is to explore the general law of education. It was carried out through educational experimentations emphasizing the control of the experimental conditions. Starting from 1990s, action research in China followed the positivist model of hypothesis verification on the one hand, while on the other hand, it introduced the notion of hermeneutics: finding and solving problems in real teaching situations and thereby understanding the real situation. Entering the new millennium, action research in China has further followed hermeneutics, accompanied by the emergence of "grassroots" action research.

Bifurcation analysis of a single species reaction-diffusion model with nonlocal delay

Jun Zhou 대한수학회 2020 대한수학회지 Vol.57 No.1

A reaction-diffusion model with spatiotemporal delay modeling the dynamical behavior of a single species is investigated. The parameter regions for the local stability, global stability and instability of the unique positive constant steady state solution are derived. The conditions of the occurrence of Turing (diffusion-driven) instability are obtained. The existence of time-periodic solutions, the existence and nonexistence of nonconstant positive steady state solutions are proved by bifurcation method and energy method. Numerical simulations are presented to verify and illustrate the theoretical results.

Pattern formation in a general Degn-Harrison reaction model

Jun Zhou 대한수학회 2017 대한수학회보 Vol.54 No.2

In this paper, we study the pattern formation to a general Degn-Harrison reaction model. We show Turing instability happens by analyzing the stability of the unique positive equilibrium with respect to the PDE model and the corresponding ODE model, which indicate the existence of the non-constant steady state solutions. We also show the existence periodic solutions of the PDE model and the ODE model by using Hopf bifurcation theory. Numerical simulations are presented to verify and illustrate the theoretical results.

The influence of bone graft procedures on primary stability and bone change of implants placed in fresh extraction sockets

Jun, Sang Ho,Park, Chang-Joo,Hwang, Suk-Hyun,Lee, Youn Ki,Zhou, Cong,Jang, Hyon-Seok,Ryu, Jae-Jun Korean Association of Maxillofacial Plastic and Re 2018 Maxillofacial Plastic Reconstructive Surgery Vol.40 No.-

Background: This study was to evaluate the effect of bone graft procedure on the primary stability of implants installed in fresh sockets and assess the vertical alteration of peri-implant bone radiographically. Methods: Twenty-three implants were inserted in 18 patients immediately after tooth extraction. The horizontal gap between the implant and bony walls of the extraction socket was grafted with xenografts. The implant stability before and after graft procedure was measured by Osstell Mentor as implant stability quotient before bone graft (ISQ bbg) and implant stability quotient after bone graft (ISQ abg). Peri-apical radiographs were taken to measure peri-implant bone change immediately after implant surgery and 12 months after implant placement. Data were analyzed by independent t test; the relationships between stability parameters (insertion torque value (ITV), ISQ abg, and ISQ bbg) and peri-implant bone changes were analyzed according to Pearson correlation coefficients. Results: The increase of ISQ in low primary stability group (LPSG) was 6.87 ± 3.62, which was significantly higher than the increase in high primary stability group (HPSG). A significant correlation between ITV and ISQ bbg (R = 0.606, P = 0.002) was found; however, age and peri-implant bone change were not found significantly related to implant stability parameters. It was presented that there were no significant peri-implant bone changes at 1 year after bone graft surgery. Conclusions: Bone graft procedure is beneficial for increasing the primary stability of immediately placed implants, especially when the ISQ of implants is below 65 and that bone grafts have some effects on peri-implant bone maintenance.