Biotechnology and Applied Biochemistry

Cover image for Vol. 61 Issue 3

Edited By: G. Gilardi and J.J. Zhong

Online ISSN: 1470-8744

Associated Title(s): Biochemistry and Molecular Biology Education, BioFactors, IUBMB Life

Featured

  • Study of the effect of F17A mutation on characteristics of Bacillus thermocatenulatus lipase expressed in Pichia pastoris using in silico and experimental methods

    Study of the effect of F17A mutation on characteristics of Bacillus thermocatenulatus lipase expressed in Pichia pastoris using in silico and experimental methods

    The position of Gln 185 and Phe-17 in the active site of closed conformation in (native BTL2) model prepared by MODELLER. There is no hydrogen bond between those two residues.

  • Continuous ethanol production from sugarcane molasses using a newly designed combined bioreactor system by immobilized Saccharomyces cerevisiae

    Continuous ethanol production from sugarcane molasses using a newly designed combined bioreactor system by immobilized Saccharomyces cerevisiae

    Bioreactor system for molasses continuous ethanol fermentation. 1: Fresh media, 2: peristaltic pump, T3: a tubular bioreactor packed with PVA immobilized yeast gel, T4: a tubular bioreactor packed with sugarcane pieces, T5: a tubular bioreactor packed with spiral wound cotton towel, 6: a broth storage tank, 7: an air pump, 8: an exhaust gas washing tank, and 9: a valve.

  • Cloning and in vitro function analysis of codon-optimized FatI gene

    Cloning and in vitro function analysis of codon‐optimized FatI gene

    Boer goat fibroblasts transfected with pEGFP-N1/AseI. F2 Boer goat fibroblasts were transfected with pEGFP-N1/AseI by electroporation as control. Images were visualized after 24 H with white light (A) and blue light (B), respectively. The transfection efficiency was about 80%, which was calculated by cell counts with green fluorescence divided by cell counts with white light. Positive clones were detected by genomic DNA PCR (C; 1,4,5 were positive) and Southern blot (D). The sequence of PCR product with genomic DNA as template showed identification with the optimized sequence (E).

  • Effect of uracil addition on proteomic profiles and 1,3-β-glucan production in Agrobacterium sp.

    Effect of uracil addition on proteomic profiles and 1,3‐β‐glucan production in Agrobacterium sp.

    2D silver-stained SDS-PAGE images of Agrobacterium proteins. (a) Growth (at 22 H, cell growth phase). (b) Production (at 90 H, before uracil addition), and (c) uracil (at 94 H, after uracil addition). Samples were taken at 22, 90, and 94 H. Uracil was added at 90 H. Three biological replicate gels were scanned, averaged using imaging software, and compared by spot matching and spot densitometry.

  • Effect of membranes on oxygen transfer rate and consumption within a newly developed three-compartment bioartificial liver device: Advanced experimental and theoretical studies

    Effect of membranes on oxygen transfer rate and consumption within a newly developed three‐compartment bioartificial liver device: Advanced experimental and theoretical studies

    (a) Schematic drawing of the commercially available bioreactor. (b) Schematic drawing of the newly developed FIF bioreactor.

  • Development of an immuno-based colorimetric assay for white spot syndrome virus

    Development of an immuno‐based colorimetric assay for white spot syndrome virus

    Schematic illustration of the immobilization and protein interaction.

  • Immunohistochemical characterization of novel murine monoclonal antibodies against human placenta-specific 1

    Immunohistochemical characterization of novel murine monoclonal antibodies against human placenta‐specific 1

    Assessment of peptide conjugation. BSA (lane 2), BSA-linker (lane 3), and BSA-conjugated peptide (lane 4) were electrophoresed on SDS-PAGE gel to assess conjugation efficacy. The smear pattern of electrophoresis movement of conjugate shows successful conjugation. Lane 1: molecular weight markers.

  • Study of the effect of F17A mutation on characteristics of Bacillus thermocatenulatus lipase expressed in Pichia pastoris using in silico and experimental methods
  • Continuous ethanol production from sugarcane molasses using a newly designed combined bioreactor system by immobilized Saccharomyces cerevisiae
  • Cloning and in vitro function analysis of codon‐optimized FatI gene
  • Effect of uracil addition on proteomic profiles and 1,3‐β‐glucan production in Agrobacterium sp.
  • Effect of membranes on oxygen transfer rate and consumption within a newly developed three‐compartment bioartificial liver device: Advanced experimental and theoretical studies
  • Development of an immuno‐based colorimetric assay for white spot syndrome virus
  • Immunohistochemical characterization of novel murine monoclonal antibodies against human placenta‐specific 1

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Engineering the MEP pathway enhanced ajmalicine biosynthesis
Kai Chang1, Fei Qiu, Min Chen, Lingjiang Zeng, Xiaoqiang Liu, Chunxian Yang, Xiaozhong Lan, Qiang Wang, and Zhihua Liao.
Volume 61, Issue 3

The terpenoid indole alkaloids (TIAs) consisting of the terpenoid moiety and the indole moiety include many high-value-added pharmaceutical alkaloids such as ajmalicine, vinblastine, and vincristine. The terpenoid moiety is provided by the plastidial 2-C-methyl-D-erythritol-4-phosphate (MEP) pathway (nonmevalonate pathway) and the indole moiety by the tryptamine pathway. Catharanthus roseus is the model plant to unveil biosynthesis of TIAs at the molecular and biochemical levels. Up to now, the reported metabolic engineering of TIA biosynthesis mainly focused on genetic modification of the indole moiety biosynthetic pathway and the strictosidine-after pathway. This study did the genetic modification of the MEP pathway by overexpressing DXR and MECS in hairy root cultures of C. roseus, and the effect of overexpressing STR combined with DXR and MECS was also discussed. The results indicated that the transgenic hairy root cultures with co-overexpression of DXR and STR or MECS and STR had higher levels of ajmalicine than those with overexpression of a single gene alone such as DXR, MECS, and STR.

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