Advances in Applied Biotechnology

Proceedings of the 3rd International

Conference on Applied Biotechnology
ICAB2016 November 25 27 2016 Tianjin
China 1st Edition Hao Liu
Visit to download the full and correct content document:
https://textbookfull.com/product/advances-in-applied-biotechnology-proceedings-of-th
e-3rd-international-conference-on-applied-biotechnology-icab2016-november-25-27-2
016-tianjin-china-1st-edition-hao-liu/
More products digital (pdf, epub, mobi) instant
download maybe you interests ...

Advances in Applied Economic Research Proceedings of

the 2016 International Conference on Applied Economics
ICOAE 1st Edition Nicholas Tsounis

https://textbookfull.com/product/advances-in-applied-economic-
research-proceedings-of-the-2016-international-conference-on-
applied-economics-icoae-1st-edition-nicholas-tsounis/

Experimental and Applied Mechanics, Volume 4:

Proceedings of the 2016 Annual Conference on
Experimental and Applied Mechanics 1st Edition Yong Zhu

https://textbookfull.com/product/experimental-and-applied-
mechanics-volume-4-proceedings-of-the-2016-annual-conference-on-
experimental-and-applied-mechanics-1st-edition-yong-zhu/

Advances in Time Series Data Methods in Applied

Economic Research International Conference on Applied
Economics ICOAE 2018 Nicholas Tsounis

https://textbookfull.com/product/advances-in-time-series-data-
methods-in-applied-economic-research-international-conference-on-
applied-economics-icoae-2018-nicholas-tsounis/

Biotechnology and Biological Sciences Proceedings of

the 3rd International Conference of Biotechnology and
Biological Sciences BIOSPECTRUM 2019 August 8 10 2019
Kolkata India 1st Edition Ramkrishna Sen (Editor)
https://textbookfull.com/product/biotechnology-and-biological-
sciences-proceedings-of-the-3rd-international-conference-of-
biotechnology-and-biological-sciences-
Advances in Speech and Language Technologies for
Iberian Languages Third International Conference
IberSPEECH 2016 Lisbon Portugal November 23 25 2016
Proceedings 1st Edition Alberto Abad
https://textbookfull.com/product/advances-in-speech-and-language-
technologies-for-iberian-languages-third-international-
conference-iberspeech-2016-lisbon-portugal-
november-23-25-2016-proceedings-1st-edition-alberto-abad/

Enhancing business stability through collaboration :

proceedings of the International Conference on Business
and Management Research (ICBMR 2016), October 25-27,
2016, Lombok, Indonesia 1st Edition Ari Kuncoro
https://textbookfull.com/product/enhancing-business-stability-
through-collaboration-proceedings-of-the-international-
conference-on-business-and-management-research-
icbmr-2016-october-25-27-2016-lombok-indonesia-1st-edition-ar/

Advances in Systems Science: Proceedings of the

International Conference on Systems Science 2016 (ICSS
2016) 1st Edition Jerzy ■wi■tek

https://textbookfull.com/product/advances-in-systems-science-
proceedings-of-the-international-conference-on-systems-
science-2016-icss-2016-1st-edition-jerzy-swiatek/

Advances in Biomolecular Medicine: Proceedings of the

4th BIBMC (Bandung International Biomolecular Medicine
Conference) 2016 and the 2nd ACMM (ASEAN Congress on
Medical Biotechnology and Molecular Biosciences),
October 4-6, 2016, Bandung, West Java, Indo 1st Edition
https://textbookfull.com/product/advances-in-biomolecular-
medicine-proceedings-of-the-4th-bibmc-bandung-international-
Fucharoen
biomolecular-medicine-conference-2016-and-the-2nd-acmm-asean-
congress-on-medical-biotechnology-and-molecular-bi/

Applied Informatics First International Conference ICAI

2018 Bogotá Colombia November 1 3 2018 Proceedings
Hector Florez

https://textbookfull.com/product/applied-informatics-first-
international-conference-icai-2018-bogota-colombia-
november-1-3-2018-proceedings-hector-florez/
Lecture Notes in Electrical Engineering 444

Hao Liu
Cunjiang Song
Arthur Ram
Editors

Advances
in Applied
Biotechnology
Proceedings of the 3rd International
Conference on Applied Biotechnology
(ICAB2016), November 25–27, 2016,
Tianjin, China
Lecture Notes in Electrical Engineering

Volume 444

Board of Series editors

Leopoldo Angrisani, Napoli, Italy
Marco Arteaga, Coyoacán, México
Samarjit Chakraborty, München, Germany
Jiming Chen, Hangzhou, P.R. China
Tan Kay Chen, Singapore, Singapore
Rüdiger Dillmann, Karlsruhe, Germany
Haibin Duan, Beijing, China
Gianluigi Ferrari, Parma, Italy
Manuel Ferre, Madrid, Spain
Sandra Hirche, München, Germany
Faryar Jabbari, Irvine, USA
Janusz Kacprzyk, Warsaw, Poland
Alaa Khamis, New Cairo City, Egypt
Torsten Kroeger, Stanford, USA
Tan Cher Ming, Singapore, Singapore
Wolfgang Minker, Ulm, Germany
Pradeep Misra, Dayton, USA
Sebastian Möller, Berlin, Germany
Subhas Mukhopadyay, Palmerston, New Zealand
Cun-Zheng Ning, Tempe, USA
Toyoaki Nishida, Sakyo-ku, Japan
Bijaya Ketan Panigrahi, New Delhi, India
Federica Pascucci, Roma, Italy
Tariq Samad, Minneapolis, USA
Gan Woon Seng, Nanyang Avenue, Singapore
Germano Veiga, Porto, Portugal
Haitao Wu, Beijing, China
Junjie James Zhang, Charlotte, USA
“Lecture Notes in Electrical Engineering (LNEE)” is a book series which reports
the latest research and developments in Electrical Engineering, namely:
• Communication, Networks, and Information Theory
• Computer Engineering
• Signal, Image, Speech and Information Processing
• Circuits and Systems
• Bioengineering
LNEE publishes authored monographs and contributed volumes which present
cutting edge research information as well as new perspectives on classical fields,
while maintaining Springer’s high standards of academic excellence. Also
considered for publication are lecture materials, proceedings, and other related
materials of exceptionally high quality and interest. The subject matter should be
original and timely, reporting the latest research and developments in all areas of
electrical engineering.
The audience for the books in LNEE consists of advanced level students,
researchers, and industry professionals working at the forefront of their fields. Much
like Springer’s other Lecture Notes series, LNEE will be distributed through
Springer’s print and electronic publishing channels.

More information about this series at http://www.springer.com/series/7818

Hao Liu Cunjiang Song
•

Arthur Ram
Editors

Advances in Applied
Biotechnology
Proceedings of the 3rd International
Conference on Applied Biotechnology
(ICAB2016), November 25–27, 2016, Tianjin,
China

123
Editors
Hao Liu Arthur Ram
College of Biotechnology Institute of Biology Leiden
Tianjin University of Science Leiden University
and Technology Leiden
Tianjin The Netherlands
China

Cunjiang Song
College of Life Sciences
Nankai University
Tianjin
China

ISSN 1876-1100 ISSN 1876-1119 (electronic)

Lecture Notes in Electrical Engineering
ISBN 978-981-10-4800-5 ISBN 978-981-10-4801-2 (eBook)
https://doi.org/10.1007/978-981-10-4801-2
Library of Congress Control Number: 2017940228

© Springer Nature Singapore Pte Ltd. 2018

This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part
of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations,
recitation, broadcasting, reproduction on microfilms or in any other physical way, and transmission
or information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar
methodology now known or hereafter developed.
The use of general descriptive names, registered names, trademarks, service marks, etc. in this
publication does not imply, even in the absence of a specific statement, that such names are exempt from
the relevant protective laws and regulations and therefore free for general use.
The publisher, the authors and the editors are safe to assume that the advice and information in this
book are believed to be true and accurate at the date of publication. Neither the publisher nor the
authors or the editors give a warranty, express or implied, with respect to the material contained herein or
for any errors or omissions that may have been made. The publisher remains neutral with regard to
jurisdictional claims in published maps and institutional affiliations.

Printed on acid-free paper

This Springer imprint is published by Springer Nature

The registered company is Springer Nature Singapore Pte Ltd.
The registered company address is: 152 Beach Road, #21-01/04 Gateway East, Singapore 189721, Singapore
Contents

Part I Microbial Genetics and Breeding

Isolation and Characterization of a Virulent Phage H6 Infecting
Lactobacillus brevis from the Fermented Chinese Cabbage . . . . . . . . . . . 3
Kechong Huang, Tian Zhang and Hongjiang Yang
Molecular Cloning and Biochemical Characterization of Oligo-1,
6-Glucosidases from Bacillus subtilis and Bacillus licheniformis . . . . . . . 13
Xiaoming Hao, Ke Cai and Zixing Dong
Transcriptomic Analysis of CYP Genes in Rhizopus nigricans
and Identification of the Steroid 11a-Hydroxylase Candidate Genes. . . . 21
Jianguo Zhao, Pengcheng Sui, Ruijie Wang, Ziyin Zhang, Fuping Lu,
Zhengxiang Wang and Xiaoguang Liu
Identification of Genes Encoding Receptors for Six
Pseudomonas aeruginosa Phages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
Jiajia You, Xiaoli Cui, Li Sun, Xiaojing Yang and Hongjiang Yang
High Efficiency Expression of Trehalose Synthase in Escherichia
coli and Its Use in the Production of Trehalose . . . . . . . . . . . . . . . . . . . . 41
Hong-Ling Liu, Rui-Ming Wang and Teng-Fei Wang
Functional Modification of the Substrate-Binding Site for Isomaltulose
Production Based on Predicted Structure of Sucrose Isomerase
from Pantoea dispersa UQ68 J . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
Huijie Liu, Xueyan Xing, Fuping Lu and Yu Li
Construction of the Escherichia coli-Bacillus subtilis Shuttle
Vector pBE2R and Identification of the Critical Residues Involved
in the Autoprocessing of the Propeptide of the Alkaline Protease . . . . . . 69
Run Wei, Xiao-mei Liang, Mei-juan Xuan, Hao-yu Yuan, Fu-ping Lu
and Ming Li

v
vi Contents

Effects of ATF2 Overexpression with BAT2 Deletion

on the Higher Alcohols and Esters in Beer Yeast . . . . . . . . . . . . . . . . . . . 79
Xiaoer Liu, Jia Xu, Li Pi, Cuiying Zhang and Dongguang Xiao
Study on a Staphylococcal Tat Signal Peptide Guided EGFP
Translocation in E. coli . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89
Qiu-Xiang Zhou, Jun Zhang, Mei-Na Wang, Wen-Hao Yang,
Jian Zhang and Qiang Gao
A Novel GH10 Xylanase Xyn13-3 from Alkaline Soil:
Gene Cloning and Heterogenous Expression . . . . . . . . . . . . . . . . . . . . . . . 97
Haiyan Qiu, Zhongyuan Li, Hui Wang, Shuang Li and Tongcun Zhang
Exploitation of a KU70-Deficient Mutant for Improving
Gene Deletion Frequency in Aspergillus niger . . . . . . . . . . . . . . . . . . . . . . 105
Liu-hua Yin, Lan Zhang, Ling Liu, Hongfei Zhang, Li Hou
and De-pei Wang
Biotransformation to Produce Boldenone by Pichia
pastoris GS115 Engineered Recombinant Strains . . . . . . . . . . . . . . . . . . . 117
Rui Tang, Peilin Ji, Ying Yu, Xu Yang, Mengjiao Liu, Kaiyuan Chen,
Yanbing Shen and Min Wang
The Prokaryotic Expression of Cyclin-Dependent Kinase 2,
and the Establish of Its Inhibitor Screening System . . . . . . . . . . . . . . . . . 125
Yuan Yuan, Meile Gao, Huan Liu, Tingting Ruan, Weiran Xie,
Meng Wu, Xin Qu, Zhen Liu, Peng Yu and Yuou Teng
Effects of Heterologous Pyruvate Carboxylase Expression
on Synthesis of L-Threonine in Escherichia coli . . . . . . . . . . . . . . . . . . . . 133
Junpeng Wang, Yan Zhao, Tao Liu, Ting Wang, Chao Han,
Qian Mao, Ning Chen and Yanjun Li
A Crystal Structure of the R-Amine Transaminase from Arthrobacter
for Asymmetric Catalysis of Chiral Amines . . . . . . . . . . . . . . . . . . . . . . . 145
Lijun Guan, Nina Ji, Qinghong Meng, Xuejun Xie, Ye Zhou,
Yijuan Cui and Shuwen Lu

Part II Optimization and Control of Biological Process

Optimization of Culture Medium for Rifamycin SV Production
by Amycolatopsis kentuckyensis 22-187 . . . . . . . . . . . . . . . . . . . . . . . . . . . 155
Mingyu Xu, Meng Li, Ying Zhang and Huitu Zhang
Optimization of Processing Parameters of Stabilizers After
Enzymes Hydrolysis for Cloudy Ginkgo Juice . . . . . . . . . . . . . . . . . . . . . 165
Haifeng Yu, Junyan Liu and Jingxi Yang
Contents vii

Characterization of Recombinant Dioxygenase

from Bacillus thuringiensis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 175
Ning Xue, Zhixiang Li, Lei Zhao, Jie Ma, Qingyang Xu,
Chenglin Zhang and Ning Chen
Optimization of Condition for Recombinant L-Isoleucine
Dioxygenase Expression in Escherich coli BL21(DE3) . . . . . . . . . . . . . . . 183
Jie Ma, Zhixiang Li, Lei Zhao, Qingyang Xu, Chenlin Zhang,
Xixian Xie and Chen Ning
Protein Sparing Effect of Carbohydrate on Growth Performance,
Digestion Ability of Common Carp (Cyprinus carpio) at Different
Feeding Frequencies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191
Ze Fan, Xiuting Qiao, Jinhui Sun, Pei Cui, Zhenzhen Fang,
Dongqing Bai and Zhenyan Cheng
Engineering of Industrial Aspergillus ochraceus Strains
for Improved Steroid 11a-Hydroxylation Efficiency via
Overexpression of the 11a-Hydroxylase Gene CYP68J5 . . . . . . . . . . . . . . 203
Xingwei Yang, Fan Wu, Xiangjiang Hou, Benfeng Lin, Ruijie Wang,
Fuping Lu, Zhengxiang Wang, Bin Zeng and Xiaoguang Liu
Response Surface Methodology Optimization Extraction
of Polysaccharides from Maca (Lepidium meyenii) Leaves
and Primary Characterization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213
Caicai Kang, Liming Zhang, Limin Hao, Huanhuan Ge,
Meng Xu, Jie Cao, Jianyong Yu and Yongwu Yang
Metabolomic Analysis of Dunaliella salina upon Concurrent
Deprivation of Nitrogen and Phosphor . . . . . . . . . . . . . . . . . . . . . . . . . . . 225
Hexin Lv, Xianggan Cui, Shilei Wang and Shiru Jia
Effect of Yeast Extract on Production of ɛ-poly-L-lysine
by Streptomyces diastatochromogenes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 235
Fengzhu Guo, Haoran Zheng, Xue Zhang, Yawen Cheng,
Zhilei Tan and Shiru Jia
Comparison of Aroma Compounds in Sauce-Flavor
and Sesame-Flavor “Shan Zhuang Lao Jiu” Liquors
by Headspace Solid-Phase Microextraction Coupled
with Gas Chromatography-Mass Spectrometry . . . . . . . . . . . . . . . . . . . . 245
Fei Liu, Wanqiang Yin, Liping Du and Dongguang Xiao
Optimization the Fermentation Conditions of Marasmius
androsaceus by Desirability Function Method. . . . . . . . . . . . . . . . . . . . . . 255
Jia Song, Le Cui, Xiaobo Ma, Yan Su, Zhengmei Huang and Min Wang
viii Contents

Heterologous Expression and Enzyme Properties of b-mannanase

from Trichoderma reesei in Pichia pastoris . . . . . . . . . . . . . . . . . . . . . . . . 267
Qing Ma, Lijuan Ma, Rui Cai, Fengchao Jiang, Pan Song
and Dongguang Xiao
Reduction of Characteristic Biogenic Amines Production
by Synergistic Fermentation of Salt-Tolerant Yeast in Soy Sauce . . . . . . 277
Wei Qi, Wen-Tao Zhang and Fu-Ping Lu
Effect of Temperature, NaCl and Ferulic Acid Concentration
on Bioconversion of Ferulic Acid to 4-Vinylguaiacol
and 4-Ethylguaiacol by Halotolerant Yeasts Candida versatilis . . . . . . . . 289
Wei Qi, Wen-Tao Zhang and Fu-Ping Lu
Near Infrared Spectroscopic (NIRS) Analysis of Polysaccharides
and Ergosterol Contents in Tricholoma matsutake Mycelium
by Improved Chemometric Model . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 299
Qiubo Chu, Quan Li, Shuang Hu, Xue Jiang, Yanzhen Wang,
Hao Zeng, Lesheng Teng and Di Wang
Antioxidant Activity and Hepatoprotective Activity of Shanxi
Aged Vinegar in Hydrogen Peroxide-Treated HepG-2 Cells . . . . . . . . . . 309
Ting Xia, Jiahui Yao, Jiankang Wang, Jin Zhang and Min Wang
Analysis and Application of the Promotion Action of Methanol
on Sorangium cellulosum Produce Epothilones . . . . . . . . . . . . . . . . . . . . . 317
Lin-hao Zhang, Lin Zhao, Xin Sun and Xin-li Liu
Optimization of Solid Fermentation Process of Bacillus megaterium
and Its Application in Crop Growth . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 329
Jinzhao Liu, Lin Zhao, Dong Ma, Xin Sun and Xin-li Liu
Comparative Quantitative Analysis of Probiotic Bacteria
During Puer Tea Pile Fermentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 339
Shuang Li, Zhongyuan Li, Cuixia Feng and Tongcun Zhang
Prediction of Hot Spots in Dimer Interface of Green Fluorescent
Protein . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 349
Wenjuan Zhang, Lin Wang, Zhiwei Sun, Bianqiang Zhang,
Qiaoqiao Tang and Qiang Gao
Regulation of NAD (H) Pool by Overexpression of Nicotinic
Acid Phosphoribosyltransferase for AD (D) Production
in Mycobacterium neoaurum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 357
Liqiu Su, Yanbing Shen, Tian Gao, Le Cui, Jianmei Luo and Min Wang
Study on the Different Replacing Groups of Trans-Paroxol
for Enzymatic Resolution Using Molecular Simulations . . . . . . . . . . . . . . 365
Chuan Zhang, Yigang Jia, Chao Xu, He Huang and Yi Hu
Contents ix

Relationship Between Coenzyme Q10 Synthesis and Cytochrome

Accumulation in Rhodobacter sphaeroides 2.4.1 . . . . . . . . . . . . . . . . . . . . 375
Penghao Li, Dan Gao, Junqian Gao, Hao Liu and Zhengliang Qi
Optimization of the One-Step and Two-Step Transformation
Methods of Mannitol by Lactobacillus buchneri . . . . . . . . . . . . . . . . . . . . 385
Hongbin Wang, Yu Li, Yongshuai Wang, Yan Chen, Yong Zhang,
Yongrui Feng, Haichao Han, Shuqi Gui and Fuping Lu
Optimization of Electroporation Conditions for Arthrobacter
simplex . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 393
Jianmei Luo, Haijie Xue, Shengnan Zhao, Xiangsheng Liu,
Xiangrong Chen, Jiajia Liu, Yanbing Shen and Min Wang
Medium Optimization for c-Aminobutyric Acid Production
by Response Surface Methodology. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 403
Chuan-You Chang, Shen-Xi Ma, Jun Zhang and Qiang Gao
A Concise and Diversity-Oriented Strategy for the Synthesis
of Substituted 2-Pyrrolidinones via an Ugi/Intramolecular
Inverse Electron Demand Diels-Alder Sequence . . . . . . . . . . . . . . . . . . . . 413
Yan Liu, Tianyi Shang, Zhong Chen, Yaozhou Zhang, Wei Sun,
Junjie Yang and Yan Pan
Optimization of Medium Components for the Production
of Flavonoids and Soluble Protein with Phellinus igniarius
in Liquid Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 421
Yingchao Wang, Zhouli Yuan, Xinrong Tan, Zaohong Ran
and Hong Jin
Application of Sun Light Conversion Film on the Outdoor
Culture of Nostoc flagelliforme . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 433
Shigang Shen, Peipei Han, Shunyu Yao, Rongjun Guo and Shiru Jia
Determination of Ginkgolic Acids in Ginkgo Seeds Using HPLC-MS
in the Presence of Lipids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 441
Yanying Hu, Guojuan Sun, Huitu Zhang, Liming Zhang,
Tongcun Zhang and Yujie Dai
High Concentration Vinegar Production by Acetic Acid Bacteria
Using Edible Alcohol as Feedstock . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 451
Xiao-Yan Yin, Rong Zhai, Wu-Kun Zhong, Jiao Huo
and Zhong-Hua Yang
Optimization of Process for Optical Resolution of DL-Pantolactone
Using Immobilized D-Lactonohydrolase . . . . . . . . . . . . . . . . . . . . . . . . . . . 459
Mei-juan Xuan, Jian-zhong Zhang, Hao-yu Yuan, Run Wei, Fu-ping Lu
and Ming Li
x Contents

Enzymatic Synthesis of L-Cysteine by Escherichia coli

Whole-Cell Biocatalyst . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 469
Mingli Ma, Tao Liu, Heyun Wu, Fangqing Yan, Ning Chen
and Xixian Xie
Analysis on Acid, Bile, and Heat Tolerance of Probiotics
Strains in Maca-Probiotics Granule . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 479
Ya-Xin Jiang, Qing-Qing Dong, Ji-Ping Qu, Tong-Cun Zhang,
Ya-Jian Song, Zhong-Yuan Li and Xue-Gang Luo
Near-Infrared Spectroscopy for the Monitoring of Leucine
Fermentation in Corynebacterium glutamicum . . . . . . . . . . . . . . . . . . . . . 487
Hongbo Wei, Tuo Shi, Lihong Du, Qixin Chen, Yuechao Ma,
Quanwei Zhang, Qian Ma and Ning Chen
Modification of Corynebacterium glutamicum YILW
for Isoleucine Production Improvement . . . . . . . . . . . . . . . . . . . . . . . . . . 495
Ning Xue, Zhixiang Li, Junjie Zhan, Jie Ma, Qingyang Xu,
Chenglin Zhang and Ning Chen
Liquid-Liquid Extraction of Hydrocortisone from the Fermentation
Liquor Contain Hydroxropyl-b-Cyclodextrin . . . . . . . . . . . . . . . . . . . . . . 505
Dongchao Yuan, Yanling Dong, Yanbing Shen, Qing Zhao and Min Wang
Conversion of Food Waste and Feldspar into Biofertilizer Using
a Stress-Tolerant Keldspar-Solubilizing Bacillus Subtilis
Xue-113168 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 513
Shengping Xue, Liangtian Miao, Jinjun Xue, Hanbo Yan
and Guiqin Li

Part III Biological Separation and Biological Purification

Biological Degradation of Aflatoxin B1 by Emericellopsis sp. 1912
and Sarocladium sp. 10A . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 525
Hui Wang, Zhongyuan Li, Haiyan Qiu, Shuang Li and Tongcun Zhang
Study on Ultrasonic-Assisted Extraction of Star Anise Oleoresin
from the Fruits of Illicium verum and Preliminary Investigation
of Its Antimicrobial Activity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 533
Ping Li, Zhan Shu, Lan Zhang, Tao Li and Lin Tian
Preparation of Dialdehyde Cellulose and Its Antibacterial Activity . . . . 545
Huanhuan Ge, Liming Zhang, Meng Xu, Jie Cao and Caicai Kang
Optimization of Microwave-Assisted Extraction of Total
Flavonoids from China-Hemp Leaves and Evaluation
of Its Antioxidant Activities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 555
Jie Cao, Limin Hao, Liming Zhang, Meng Xu, Huanhuan Ge, Caicai Kang,
Jianyong Yu and Zongzhen Wang
Contents xi

Comparison of Different Protocols of Gradient Ammonium Sulfate

Fractionation of Antibacterial Proteins/Peptides from Clarias
gariepinus Wastes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 569
Xiaomei Wang, Yan Wang, Yunxia Xu, Zhuanzhuan Li, Chengxun Chen,
Jinwei Gao and Tao Li
Screening of Microbial with the Ability of Epothilones
Biotransformation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 579
Meng Zhang, Lin Zhao, Xin Sun and Xin-li Liu
Isolation, Screening and Evaluation of Potential Biocontrol
Endophytes Against Ralstonia solanacearum on Ginger . . . . . . . . . . . . . . 587
Ning Zhou, Lin Zhao, Xin Sun and Xin-li Liu
Isolation and Molecular Identification of Lactobacillus brevis
from Spoilage Craft Beer in China . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 597
Zhu Liping
Production of 5′-Inosinic Acid by Whole-Cell Biocatalyst Expressing
a Mutated Acid Phosphatase/Phosphotransferase . . . . . . . . . . . . . . . . . . . 605
Hui Yuan, Zi-fan Jia, Ju-hua He, Xiao-guang Fan and Ning Chen
Bio-production of L-rhamnonate by Pseudomonas taetrolens. . . . . . . . . . 615
Shuhong Mao, Jianlin Wu, Lixia Zhang, Shuqi Gui and Fuping Lu
Preparation and Stability Evaluation of Direct Vat Set Lactobacillus
helveticus AAF1-5 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 623
Chuanxue Fu, Shuai Yang, Jun Mou, Jia Song, Min Wang and Yu Zheng
Overexpression of the Endo-inulinase Gene from Two Different
Sources and Characteristics Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . 631
Li-Kun Wei, Qing-Long Xin, Zhi-Mei Feng, Xue-Yan Xing, Wei Feng
and Yu Li
Production of Ethyl Acetate Catalyzed by Activated Carbon-Based
Solid Acid Catalyst . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 643
Jia Li, Yan Li and Hua Zhao
The Preparation of Chitosan from Corncob Hydrolyzate
by Actinomucor elegans . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 653
Yan Li, Jia Li and Hua Zhao
High-Efficiency Separation and Purification of Taq DNA Polymerase . . . 663
Hao Zhou, Yujie Zhang, Zhiyin Hu, Ai Mu and Xiangchao Gu
Determination of Activity and Extraction of Thrombin
from the Porcine Blood. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 673
Tianjun Li, Heng Li, Hong Pan, Tao Li and Jun Shi
xii Contents

An Efficient Method for Isolation and Separation of Pigments

from Streptomyces alboflavus TD-1 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 681
Xiaoyue Gu, Yali Zhang, Laifeng Lu, Zhenjing Li and Changlu Wang

Part IV Progress of Biotechnology

Establishment of the Method for Screening Small Molecule Inhibitors
Blocking the Interaction Between PD-1 and Its Ligand PD-L1 . . . . . . . . 695
Lei Jing, Fushan Yan, Yingchun Wang, Bo Jiang, Li Chang, Cheng Cheng,
Yuyin Li and Aipo Diao
Construction of the PD-L1 Promoter-Luciferase Reporter Expressing
Vector for Small Molecule Inhibitors Screening . . . . . . . . . . . . . . . . . . . . 705
Bo Jiang, Zhichen Shi, Ali Wang, Yuyin Li, Qiurong Zhang, Lei Jing
and Aipo Diao
Construction and Functional Analysis of BNP Promoter Luciferase
Reporter Plasmid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 713
Jian Zhang, Nan Wang, Yanzhong Liu, Man Li, Huiqin Gong,
Hongpeng He and Tongcun Zhang
HPV18 E6 and E7 Influence the Expression of Cancer Related
LncRNAs in HeLa Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 719
Xiang Liu, Yongwei Lai, Hailin Yao, Mengmeng Zhang, Hao Zhou,
Tongcun Zhang and Hongpeng He
Construction of Tip60-Encoding Plasmid and the Effect of Tip60
on the Expression of HPV18 Genes in HeLa Cells . . . . . . . . . . . . . . . . . . 729
Yongwei Lai, Xuena Liu, Yijie Wang, Yunpeng Yue, Xiang Liu,
Hao Zhou, Nan Wang, Xue-Gang Luo, Wenjian Ma, Tong-Cun Zhang
and Hongpeng He
Investigation of Aquatic Pathogens and Diversity Analysis
of Aeromonas Isolates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 737
Zhaoyuan Jing, Yang He, Qian Li, Bo Zhang and Hongjiang Yang
Expression of Transcription Factor EB (TFEB) Promotes Cancer
Cell Proliferation, Migration and Invasion . . . . . . . . . . . . . . . . . . . . . . . . 745
Wei Li, Yang Liu, Min Hao, Meng Yang, Shuang Zhao, Zhenxing Liu
and Aipo Diao
Research Progress of Squalene Synthase on Function
and Application . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 755
Dengyue Sun, Qianqian Guo, Zhangliang Zhu, Songtao Li,
Jian-Wen Wang, Yu-Fu Zhang, Lijun Guan, Hui-Min Qin and Fuping Lu
Review in Metabolic Modulation of Higher Alcohols
in Top-Fermenting Yeast . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 767
Zhongguan Sun and Dongguang Xiao
Contents xiii

Research Progress of Aldehyde Ketone Reductase for Asymmetric

Catalysis of Chiral Compounds . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 775
Songtao Li, Zhangliang Zhu, Jian-Wen Wang, Qianqian Guo, Panpan Xu,
Dengyue Sun, Hui-Min Qin and Fuping Lu
Population Structure and Genetic Diversity Analysis of Peanut
(Arachis hypogaea L.) Using Molecular Markers . . . . . . . . . . . . . . . . . . . 783
Xiu Rong Zhang, Feng Zhen Liu, Kun Zhang and Yong Shan Wan
Research on c-Polyglutamic Acid Fermentation with Extract
from Waste Beer Yeast . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 795
Jun Yu, Lin Zhao, Song Li, Xin Sun and Xin-li Liu
Isolation and 16SrDNA Identification of Bacteria from Traditional
Kazak Dairy Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 803
Wenyuan Sun, Yanli Fan, Jing Li, Gaoshaer Kayierhali, Xuejiao Liu,
Yun Hao, Yirong Hou, Yajian Song and Tongcun Zhang
Research on Extracting Technology of Chlorogenic Acid
from Honeysuckle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 811
Yang Sun, Ye-Min Yu, Hong-Bo Suo, Ying-Lan Zhu, He Huang
and Yi Hu
Construction and Functional Analysis of Luciferase Reporter
Plasmid Containing Vimentin Gene Promoter . . . . . . . . . . . . . . . . . . . . . 823
Cheng-Xi Yu, Yuan Xiang, Xing-Hua Liao, Xiao-Yu Zhang, Hui Li,
Jia-Peng Li and Tong-Cun Zhang
STAT5A and MKL-1 Activate the Activity of Luciferase Reporter
Plasmid Containing FOXP3 Gene Promoter . . . . . . . . . . . . . . . . . . . . . . . 829
Jia-Peng Li, Hui Li, Xiao-Yu Zhang, Cheng-Xi Yu, Yuan Xiang, Ze Yin,
Xing-Hua Liao and Tong-Cun Zhang
Study the Role of E-selectin and Its Ligand sLeX in the Adhesion
Between THP-1 Cells and HUVEC Cells. . . . . . . . . . . . . . . . . . . . . . . . . . 839
Qian Zhang, Huan Liu, Chaoran Yao, Tingshen Li, Xuehui Li, Li Zhang,
Zhen Liu, Peng Yu and Yuou Teng
The Application of Microbial Technology in Harbor Engineering:
The Impact of Extracellular Polymeric Substances
on the Sedimentation and Properties of Fluid Mud . . . . . . . . . . . . . . . . . 847
Xiaohua Chen, Qixiu Pang, Mengnan Li, Baojiang Sun, Ruibo Zhang,
Shiru Jia and Peipei Han
Polyvalent Effect Enhances Anti-influenza Virus Activity . . . . . . . . . . . . 859
Haipeng Liu, Haojie He, Zhaoliang Yang, Peng Yu and Kui Lu
 Part I
Microbial Genetics and Breeding
Isolation and Characterization
of a Virulent Phage H6 Infecting
Lactobacillus brevis from the Fermented
Chinese Cabbage

Kechong Huang, Tian Zhang and Hongjiang Yang

1 Introduction

Nowadays, traditional vegetable fermentation products contain fermented cucum-

ber, kimchi, sauerkraut and fermented Chinese cabbage, and etc. They are prepared
by a variety of lactic acid bacteria (LAB) naturally present on surfaces of vegeta-
bles. The metabolic activities of LAB determine the quality and safety of the final
fermentation products [1]. Recent studies have analyzed the diversity and dynamics
of the microflora in several fermented vegetables, such as kimchi [2], sauerkraut [3]
and fermented cucumber [4, 5]. Phage is the most abundant biological entity living
on the earth widely distributed in nearly all natural niches. Many phage species
have been discovered in the fermented vegetable products. The presence of diverse
phages against LAB strains in vegetable fermentation processes can potentially
cause significant mortality to LAB, causing loss or damage to the productions.
Now, the phage ecology of several vegetable fermentations has been reported,
including sauerkraut fermentations [6, 7], and cucumber fermentations [8]. In
addition, the novel LAB phages against Weissella cibaria have been isolated from
the commercial kimchi fermentations [9].
Fermented Chinese cabbage is one of the most popular traditional fermented
Chinese foods, and it is a mainly LAB fermented vegetable product. In the cabbage
fermentation, Lactobacillus plantarum, Lactobacillus brevis and Lactobacillus
pentosus are detected as the dominant species, and others including Lactobacillus
acidophilus, Lactobacillus fermentum and Leuconostoc mesenteroides [10].
In this study, lactic acid bacteria and the corresponding phages were isolated
from the fermented Chinese cabbage samples collected in a village of Ji County,

K. Huang
T. Zhang
H. Yang (&)

Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education Tianjin,
Key Laboratory of Industrial Microbiology College of Biotechnology, Tianjin University
of Science and Technology, Tianjin 300457, China
e-mail: hongjiangyang@tust.edu.cn

© Springer Nature Singapore Pte Ltd. 2018 3

H. Liu et al. (eds.), Advances in Applied Biotechnology, Lecture Notes
in Electrical Engineering 444, https://doi.org/10.1007/978-981-10-4801-2_1
4 K. Huang et al.

Tianjin. The isolated phage was further characterized, including morphology, burst
size, latent time, genome size and host range. The data of this study may provide
valuable information for the prevention of phage infection in the production of the
fermented Chinese cabbage.

2 Materials and Methods

2.1 Growth Conditions

MRS medium was composed of 20 g/L glucose, 10 g/L peptone, 8 g/L beef
extract, 4 g/L yeast extract, 5 g/L sodium acetate, 2 g/L ammonium citrate, 0.2 g/L
MgSO4, 0.05 g/L MnSO4, 2 g/L KH2PO4, and 0.1% Tween-80 [11]. MRS
(pH = 5.0) and MRS (pH = 6.4) were used for lactic acid bacteria (LAB) isolation
and cultivation, respectively. Broth was used for liquid cultures, 1.5% solid agar
medium was used for bacteria plating, and 0.5% semi-solid agar medium was used
for phage plaque-forming assays. Both plates and liquid cultivations were statically
incubated at 30 °C.

2.2 Isolation and Identification of Lactic Acid Bacteria

The fermented Chinese cabbage samples were collected in Ji County, Tianjin,

China on March 16th, 2016. To isolate lactic acid bacteria, serial dilutions of nine
fresh fermented Chinese cabbage samples were spread on MRS agar. The identity
of these strains were determined by analyzing the 16S rDNA gene sequence [12].

2.3 Isolation and Propagation of Phage

The isolated LAB strains were used as indicator for isolating phage from the
fermented Chinese cabbage samples. In brief, added 1 mL samples to 100 mL
MRS medium, and incubated at 30 °C for 48 h to enrich phages. The phages were
isolated by the double-layer plating technique. The propagation of phage was done
as previously described in order to obtain high titer lysates [13]. Phage lysate was
filtered through a 0.2 lm-pore size sterile filter and stored at 4 °C for future use.
Isolation and Characterization of a Virulent Phage H6 Infecting … 5

2.4 Transmission Electron Microscopy

The phage preparations was further treated with DNase I (1 lg/mL) and RNase A
(1 lg/mL) at 37 °C for 30 min. The treated lysate was washed ten times with
0.1 mol/L ammonium acetate solution (pH = 7.0) using the 100 kD Amicon filters.
The retained phage solution was used directly for negative staining as described
previously [14]. Photographs were taken with a JROL1011 transmission electron
microscope operating at 100 kV.

2.5 Host Range Determination

As described previously [15], spotting assay was conducted to determine the sen-
sitivity of the isolated phages to the host strains. The double-layer method was used
to further confirm the above results.

2.6 Restriction Analysis of the Phage Genomic DNA

The 100 mL phage lysate was used for phage DNA extraction using the
phenol-chloroform method described previously [16]. Purified phage genomic
DNA was subjected to digestions with several restriction endonucleases, including
EcoRI, EcoRV, XbaI, HindIII, SmaI, StuI and BamHI, respectively. The genome
size was estimated by compilation of DNA fragment sizes resulting from the seven
restriction enzymes digestion profiles.

2.7 SDS-PAGE Analysis of the Phage Structural Proteins

Purified phage particles were further filtered through the Amicon-100 filter, and
washed three times with 0.1 mol/L ammonium acetate solution (pH = 7.0). Treated
phage particles were subjected to SDS-PAGE directly, and the gel stained with
Coomassie Blue R-250.

2.8 One Step Growth Curve

One-step growth experiment was carried out according to the previous descriptions
[17]. In brief, 50 mL bacterial cells were incubated to mid-exponential-phase
(OD600 = 0.5–0.6) and harvested by centrifugation at 6000 rpm for 10 min.
6 K. Huang et al.

The pellet was resuspended in 0.5 mL fresh MRS medium and mixed with 0.5 mL
phage solution (1  106 pfu/mL). Phage was allowed to adsorb for 1 min and the
mixture was subjected to centrifuge immediately at 13,000 rpm for 30 s to remove
free phage particles. The treated pellet was resuspended in 100 mL fresh MRS
medium and the culture was continuously incubated at 30 °C. Samples were taken
at 15 min intervals and phage titre was determined by the double-layer plate
method. The latent period was deduced from the triphasic curve. The burst size of
phage was calculated by dividing the phage titers at the plateau phase by the
infective centers number at the latent phase.

2.9 The Lysis Curve of Host Strain by Phage

The logarithmic cultures of the strain J68 were mixed with the phage lysate at
different MOIs (MOI = 100, 10, 1, 0.1, 0.01, 0). The 96-well plate was filled with
the 300 lL mixture per well, and the value of OD600 was measured at 2 h intervals
by ELISA reader (GS40A24).

3 Results

3.1 Identification of Lactic Acid Bacteria

Eight isolated strains from the fermented Chinese cabbage samples were confirmed
the identity by analyzing their 16S rDNA gene sequence. The resulted sequences
were deposited to GenBank and aligned to search for the most similar sequences. In
final, six collected strains were validated to be Lactobacillus brevis, other two
strains belonged to Lactobacillus plantarum.

3.2 Isolation and Titer of Phage

L. brevis J68 was used as indicator strain for virulent bacteriophages screening from
the fermented Chinese cabbage samples. A phage was eventually isolated and
named H6. Its plaques were circular, clear and transparent with smooth edge,
showing 1–2 mm in diameter. At the MOI = 0.01, titer of phage H6 reached
1  109 pfu/mL in MRS and 5  109 pfu/mL in MRS with CaCl2 (2.375 g/L).
Isolation and Characterization of a Virulent Phage H6 Infecting … 7

3.3 Host Range Analysis

The susceptibility to phage H6 was investigated with isolated strains from fer-
mented Chinese cabbage samples and other strains, they included six L. brevis, six
L. plantarum, two Lactobacillus vaginalis, nine Lactobacillus reuteri, two
Weissella cibaria, one Lactobacillus curvatus and one Lactobacillus johnsonii. We
found that all six L. brevis strains were sensitive to phage H6, and other strains all
were resistant to phage H6. The sequences of 16S rDNA showed that there were
differences among six L. brevis strains. The result indicated that phage H6 might
have a broad host range and was capable of infecting multiple isolates of
Lactobacillus brevis, however, phage H6 didn’t infect lactic acid bacteria from
other genera.

3.4 Morphology Study by Transmission Electron

Microscopy

The treated phage solution was used directly for negative staining. Images of phage
H6 were developed using transmission electron microscope (Fig. 1). The obtained
image showed that phage H6 had an icosahedra head of 93.3 nm in diameter and a
long contractile tail about 166.6 nm in length, and it was classified as a lytic phage
of Myoviridae in Caudovirales.

Fig. 1 Morphology of phage

H6 with the transmission
electron micrograph method.
The scale bar represented
100 nm
8 K. Huang et al.

3.5 Restriction Fragment Analysis of Genomic DNA

Phage H6 was amplified and its genomic DNA was extracted. Purified genomic
DNA was digested with several restriction endonucleases, including EcoRI,
EcoRV, XbaI, HindIII, SmaI, StuI and BamHI, as subsequently subjected to elec-
trophoretic analyses (Fig. 2). Based on the digestion profiles of EcoRI, EcoRV,
XbaI, and HindIII, the genome size was determined to be approximately at the
range of 59.6–61.2 kb. The restriction enzymes analysis also indicated that phage
H6 was a dsDNA virus.

3.6 Proteomic Analysis of Phage Structural Proteins

Purified phage particles were subjected to SDS-PAGE and proteomic patterns were
obtained after Coomassie Blue R-250 staining (Fig. 3). Totally, six protein bands
were displayed on the gel with the molecular weights ranging approximately from
30 to 60 kD.

Fig. 2 Restriction fragments

analysis of phage genomic
DNA. Phage genomic DNA
was digested with EcoRI
(lane 1), EcoRV (lane 2),
XbaI (lane 3), HindIII (lane
4), SmaI (lane 5), StuI (lane
6), and BamHI (lane 7),
respectively. M1: 15 kb DNA
marker 1; M2: 5 kb DNA
marker 2
Isolation and Characterization of a Virulent Phage H6 Infecting … 9

Fig. 3 SDS-PAGE of the

viral structural proteins. Solid
arrows indicated the major
proteins bands; open arrows
showed the minor proteins
bands

3.7 Latent Time and Burst Size of Phage H6

One-step growth experiment was performed to determine the latent time and burst
size of phage H6. As inferred from the triphasic curve (Fig. 4), the latent period was
about 90 min and the burst size was about 40.4 pfu/infection center.

Fig. 4 One-step growth

curve. Latent time and burst
size of phage H6 were
inferred from the triphasic
curve. L: latent phase; R: rise
phase; P: plateau phase
10 K. Huang et al.

Fig. 5 Lysis curve of host

strain by phage H6 with
different MOIs

3.8 Lysis Curve of Host Strain by Phage H6

The lysis ability of phage H6 to the Lactobacillus brevis J68 was measured at
different MOIs. As shown in Fig. 5, with the increase of phage titeres, the bacte-
riostatic ability of phage H6 gradually strengthened. At the MOI = 1, the growth of
strain J68 appeared to decline in the 10th hour; the strain J68 hardly grew at the
MOI = 10 or 100.

4 Discussion

To achieve high quality of fermented products, knowledge about the diversity,

abundance, and property of phages in vegetable fermentation products are essential
for developing phage control strategies. Lu et al. [6] investigated the ecology of
phages infecting lactic acid bacteria in commercial sauerkraut fermentations, the
result indicated that the phages against Leuconostoc, Weissella and Lactobacillus
were widely existed in sauerkraut fermentations. In one recent study, a high abun-
dance of phage DNA was found in kimchi fermentation [18]. And several LAB phages
against L. plantarum and Pediococcus sp. have been isolated from commercial
cucumber fermentations [7, 19]. Bacteriophage infection was a well-recognized
problem in industrial food fermentations and a variety of countermeasures were
employed to control the problem [20]. Therefore, the gain of more information about
bacteriophages in vegetable fermentation products was essential to the phage-control
strategies.
In this study, a virulent phage infecting L. brevis was isolated from fermented
Chinese cabbage samples, and its biological characteristics were determined. The
restriction analysis indicated that phage H6 was a dsDNA virus with an approxi-
mate genome size of 59.6–61.2 kb, but, it was uncertain that phage H6 genome was
Isolation and Characterization of a Virulent Phage H6 Infecting … 11

linear or circular. The analysis of phage H6 structural proteins showed that there
was a huge protein band at 60 kD, we speculated that it contains several protein
bands and this assumption can be verified by mass spectrometry. The data from this
study can provide more information about L. brevis bacteriophages. The objectives
of this study were to provide more formulation in order to prevent infection of
bacteriophages in fermented Chinese cabbage.

Acknowledgements This work was partly supported by The National Natural Science
Foundation of China (Grants 31370205 and 30970114).

References

1. Pederson CS, Albury MN (1969) The sauerkraut fermentation. New York State Agricultural
Experiment Station Technical Bulletin 824. Geneva, New York
2. Lee JS, Heo GY, Lee JW, Oh YJ, Park JA, Park YH, Pyun YR, Ahn JS (2005) Analysis of
kimchi microflora using denaturing gradient gel electrophoresis. Int J Food Microbiol
102(2):143–150
3. Plengvidhya V, Breidt F Jr, Lu Z, Fleming HP (2007) DNA fingerprinting of lactic acid
bacteria in sauerkraut fermentations. Appl Environ Microbiol 73(23):7697–7702
4. Medina E, Pérez-Díaz IM, Breidt F, Hayes J, Franco W, Butz N, Azcarate-Peril MA (2016)
Bacterial ecology of fermented cucumber rising pH spoilage as determined by
nonculture-based methods. J Food Sci 81(1):M121–M129
5. Chen YS, Wu HC, Lo HY, Lin WC, Hsu WH, Lin CW, Lin PY, Yanagida F (2012) Isolation
and characterisation of lactic acid bacteria from jiang-gua (fermented cucumbers), a
traditional fermented food in Taiwan. J Sci Food Agric 92(10):2069–2075
6. Lu Z, Breidt F, Plengvidhya V, Fleming HP (2003) Bacteriophage ecology in commercial
sauerkraut fermentations. Appl Environ Microbiol 69:3192–3202
7. Yoon S-S, Barrangou-Poueys R, Breidt F, Klaenhammer TR, Fleming HP (2002) Isolation
and characterization of bacteriophages from fermenting sauerkraut. Appl Environ Microbiol
68(2):973–976
8. Lu Z, Perez-Diaz IM, Hayes JS, Breidt F (2012) Bacteriophage ecology in a commercial
cucumber fermentation. Appl Environ Microbiol 78(24):8571–8578
9. Kleooen HP, Holo H, Jeon SR, Nes IF, Yoon SS (2012) Novel Podoviridae family
bacteriophage infecting Weissella cibaria isolated from Kimchi. Appl Environ Microbiol 78
(20):7299–7308
10. Yan P, Chai Z, Xue W, Chang X, Kong D, Zhang H (2009) Lactic acid bacteria diversity in
fermented cabbage estimated by culture-dependent and -independent. Wei Sheng Wu Xue
Bao 49(3):383–388
11. De Man JC, Rogosa M, Sharpe ME (1960) A medium for the cultivation of lactobacilli.
J Appl Bacteriol 23(1):130–135
12. Gürtler V, Stanisich VA (1996) New approaches to typing and identification of bacteria using
the 16S–23S rDNA spacer region. Microbiology 142(Pt 1):3–16
13. Jaomanjaka F, Ballestra P, Dols-lafargue M, Le Marrec C (2013) Expanding the diversity of
oenococcal bacteriophages: insights into a novel group based on the integrase sequence. Int J
Food 166(2):331–340
14. Nugent KM, Cole RM (1977) Characterization of group H streptococcal temperate
bacteriophage phi 227. J Virol 21(3):1061–1073
12 K. Huang et al.

15. Yang H, Liang L, Lin S, Jia S (2010) Isolation and characterization of a virulent
bacteriophage of Acinetobacter baumannii. BMC Microbiol 10:131; PMID: 20426877;
1471-2180-10-131
16. Sambrook J, Russell DW (2001) Molecular cloning. CSHL Press, New York
17. Chow JJ, Batt CA, Sinskey AJ (1988) Characterization of Lactobacillus bulgaricus
bacteriophage ch2. Appl Environ Microbiol 54(5):1138–1142
18. Jung JY, Lee SH, Kim JM, Park MS, Bae JW, Hahn Y, Madsen EL, Jeon CO (2011)
Metagenomic analysis of kimchi, a traditional Korean fermented food. Appl Environ
Microbiol 77(7):2264–2274
19. Yoon S-S, Barrangou-Poueys R, Breidt F, Fleming HP (2007) Detection and characterization
of a lytic Pediococcus bacteriophage from the fermenting cucumber brine. J Microbiol
Biotechnol 17:262–270
20. Moineau S, Levesque C (2005) Control of bacteriophages in industrial fermentation. In:
Kutte E, Sulakvelidze A (eds) Bacteriophage: biology and applications. CRC Press, Boca
Raton, pp 286–296
Molecular Cloning and Biochemical
Characterization of Oligo-1,6-Glucosidases
from Bacillus subtilis and Bacillus
licheniformis

Xiaoming Hao, Ke Cai and Zixing Dong

1 Introduction

Oligo-1,6-glucosidase (O-1,6-G, EC 3.2.1.10), belonging to the subfamily 31 of the

glycoside hydrolase family 13 (GH13_31) [1] catalyzes the exo hydrolysis of a-1,6-
glucoside bonds from the non-reducing ends of a-limit dextrin, isomaltose and
other isomaltooligosaccharides (IMOs), but has no activity towards a-1,4-glucoside
bonds of malto-oligosaccharides [2]. Acting together with maltase, oligo-1,6-glu-
cosidase can completely hydrolyze a-amylase dextrins, allowing the complete
digestion of starch in the gastrointestinal tract of mammals [3, 4]. Moreover, since
novel oligosaccharides are finding increasing applications in biotechnological and
chemical industries, the debranching enzymes containing oligo-1,6-glucosidases are
also valuable [5].
In our study, two genes encoding oligo-1,6-glucosidases from Bacillus subtilis
and B. licheniformis were cloned and overexpressed in Pichia pastoris, and their
enzymatic properties were comprehensively investigated.

X. Hao
Z. Dong (&)

Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education,
School of Biotechnology, Tianjin University of Science and Technology,
Tianjin 300457, China
e-mail: dzx@tust.edu.cn
X. Hao
K. Cai
Z. Dong
School of Chemical Engineering and Materials Science, Tianjin University
of Science and Technology, Tianjin 300457, China
X. Hao
Z. Dong
Tianjin Economic-Technological Development Area,
No. 29, the 13th Street, Tianjin, China

© Springer Nature Singapore Pte Ltd. 2018 13

H. Liu et al. (eds.), Advances in Applied Biotechnology, Lecture Notes
in Electrical Engineering 444, https://doi.org/10.1007/978-981-10-4801-2_2
14 X. Hao et al.

2 Materials and Methods

2.1 Bacterial Strains, Plasmids

Escherichia coli JM109, P. pastoris strain GS115 and vector pPIC9K were
obtained from Invitrogen (Carlasbad, CA).

2.2 Gene Clone

Gene encoding oligo-1,6-glucosidase from B. subtilis (bsog) was amplified by PCR

using the primer sets Bs-1 5′-GTAAGTGAATGGTGGAAAGAAGCTGTC-3′ and
Bs-2 5′-TGCTCTAGATCATATACTAATGCCCATCACTGCTT-3′. And primer
sets Bl-1 5′-GTAAGCCAATGGTGGAAAGAGGC-3′ and Bl-2 5′-TGCTCTAGA
TCATGATGTGTAATCCTTTGCCC-3′ were used for blog.

2.3 Enzyme Assays

The described method [6] was used to measure the activities of oligo-1,6-gluco-
sidases from B. subtilis and B. licheniformis.

2.4 pH and Temperature Dependence of Activity

pH optima of BsOG and BlOG were analyzed by incubating them for 15 min at
37 °C as described above.
The optimal reaction temperatures of BsOG and BlOG were determined at
temperatures ranging from 15 to 70 °C at pH 6.8.

2.5 Determination of Kinetic Parameters

For determination of enzyme kinetics, various amounts of pNPG were used as

substrates, and the activities of BsOG and BlOG were measured. Isomaltose and
isomaltotriose which were dissolved in 0.2 M phosphate buffer (pH 7.0). An
Agilent 1200 HPLC system (Waldbronn, Germany) equipped with an evaporative
light-scattering detector was used for quantification and identification of various
Molecular Cloning and Biochemical Characterization … 15

oligosaccharides in the samples. Separation of oligosaccharides was carried out

using a Prevail™ Carbohydrate ES 5l column (GRACE, 4.6 mm  250 mm i.d.;
particle size, 5 lm).

2.6 Specificity Towards Natural Substrates

Isomaltose, isomaltotriose, isomaltulose, panose, maltotriose, maltose, sucrose,

amylose, amylopectin and maltodextrin used as substrates to study the specificity
towards natural substrates, were dissolved in 0.2 M phosphate buffer (pH 7.0).

2.7 Hydrolysis of Isomaltotriose and IMOs by OGs

Hydrolysis of isomaltotriose and IMOs by BsOG and BlOG were performed and
analyzed as described above except that isomaltotriose were dissolved in 0.2 M
phosphate buffer (pH 7.0) to a final concentration of 4 mM and that samples were
withdrawn at different times for HPLC analysis.

3 Results

3.1 Gene Cloning and Expression of Oligo-1,6-Glucosidases

With genomic DNA as templates, respective genes encoding oligo-1,6-glucosidases

from B. subtilis and B. licheniformis were amplified by PCR, and the fragments
obtained were 1700 bp, consistent with their theoretical sizes (bsog, 1686 bp; blog,
1707 bp). After five days of shake flask fermentation, the activities of oligo-1,6-
glucosidases of these GS115-bsog and GS115-blog were 1085 and 1037 U/mL.

3.2 Effects of Temperature on the Activity and Stability

of Recombinant Oligo-1,6-Glucosidases

Effects of temperature on the activity and stability of recombinant oligo-l,6-glu-

cosidases were examined. The temperature optima of BsOG and BlOG were 40 and
45 °C, respectively. BsOG retained 80% of its maximum activity at temperatures
ranging from 35 to 50 °C, while BlOG had 80% of its maximum activity at tem-
peratures between 40 and 50 °C. For determination of the thermostability,
16 X. Hao et al.

BsOG and BlOG were pre-incubated in 0.2 M phosphate buffer (pH 7.0) at 50 °C,
and the residual activities were measured at the indicated times. Incubation at 50 °C
for 20 min, BsOG and BlOG activity were not detected.

3.3 Effects of pH on the Activity and Stability

of Recombinant Oligo-1,6-Glucosidases

The relative activities of BsOG and BlOG at various pHs were measured with two
different buffer systems at 37 °C. The effects of pH over a range of 4.0–10.0 on the
activities of BsOG and BlOG. The optimal pH of BsOG and BlOG were 7.0 and
6.5. BsOG had a relatively broad pH optimum ranging from 6.0 to 9.5. The opti-
mum pH range of BlOG was 5.5–7.5.

3.4 Kinetic Parameters of Recombinant

Oligo-1,6-Glucosidases

Michaelis-Menten and Lineweaver-Burk plots were used to calculate the kinetic

parameters. Although both enzymes hydrolyzed pNPG, isaomaltose and isomal-
totriose, their kinetic parameters were different. BsOG catalyzed pNPG, isomaltose
and isomaltotriose with Kms of 0.1, 0.59 and 8.60 mM. BlOG showed Km values of
0.27, 0.86 and 10.8 mM for pNPG, isomaltose and isomaltotriose.

3.5 Substrate Specificity of Recombinant

Oligo-1,6-Glucosidases

The ability of BsOG and BlOG to hydrolyze various di- and maltooligosaccharides,
as well as a-glucan polymers, such as amylose and amylopectin, was determined.
As shown in Table 1, both BsOG and BlOG hydrolyzed isomaltose, isomaltotriose,
isomaltulose, panose, sucrose, amylopectin and maltodextrin, and exhibited weak
activity against amylose. However, no activity was observed toward maltose and
maltotriose. BsOG and BlOG also exhibited a-1,2-glucosidase activity on sucrose,
in accord with the substrate specificity of isomaltase from S. cerevisiae [2]. This
restricted substrate specificity indicated that these two enzymes were oligo-1,6-
glucosidases [7]. Oligo-1,6-glucosidase prefers isomaltotriose, and hyrolyzes IMOs
and dextran [7]. On the other hand, S. cerevisiae isomaltase preferentially cleaves
Another random document with
no related content on Scribd:
The Project Gutenberg eBook of Cardinal de
Richelieu
 This ebook is for the use of anyone anywhere in the United
States and most other parts of the world at no cost and with
almost no restrictions whatsoever. You may copy it, give it away
or re-use it under the terms of the Project Gutenberg License
included with this ebook or online at www.gutenberg.org. If you
are not located in the United States, you will have to check the
laws of the country where you are located before using this
eBook.

Title: Cardinal de Richelieu

Author: Eleanor C. Price

Release date: September 10, 2023 [eBook #71607]

Language: English

Original publication: London: Methuen & Co. Ltd, 1912

Credits: Turgut Dincer and the Online Distributed Proofreading

Team at https://www.pgdp.net (This file was produced
from images generously made available by The
Internet Archive)

*** START OF THE PROJECT GUTENBERG EBOOK CARDINAL

DE RICHELIEU ***
 CARDINAL DE RICHELIEU
TRIPLE PORTRAIT BY PHILIPPE DE CHAMPAGNE
 CARDINAL DE RICHELIEU
BY
ELEANOR C. PRICE
AUTHOR OF “A PRINCESS OF THE OLD WORLD”

“Il est dans l’histoire de grandes et énigmatiques figures

sur lesquelles le ‘dernier mot’ ne sera peut-être jamais dit....
Telle est, assurément, celle du Cardinal de Richelieu.”

Baron A. de Maricourt.

WITH TWELVE ILLUSTRATIONS

SECOND EDITION

METHUEN & CO. LTD.

36 ESSEX STREET W.C.
LONDON
First Published September 19th 1912
Second Edition 1912
 INTRODUCTORY NOTE

“Temerarious indeed must he appear who attempts to

comprehend in so small a space the admirable actions of a
Hero who filled the whole earth with the fame of his glory, and
who, by the wonders he worked in our own days, effaced the
most lofty and astounding deeds of Pagan demigods and
illustrious Personages of Antiquity. But what encourages me
to attempt a thing so daring is the preciousness of the
material with which I have to deal; being such that it needs
neither the workman nor his art for the heightening of its
value. So that, however little I may say of the incomparable
and inimitable actions of the great Armand de Richelieu, I
shall yet say much; knowing also that if I were to fill large
volumes, I should still say very little.”

Although the courtly language of the Sieur de la Colombière,

Gentleman-in-Ordinary to Louis XIV., who wrote a Portrait of
Cardinal de Richelieu some years after his death, may appear
extravagant to modern minds, there is no denying that he is justified
on one point—the marvellous interest of his subject.
Few harder tasks could be attempted than a complete biography
of Richelieu. It would mean the history of France for more than fifty
years, the history of Europe for more than twenty: even a fully
equipped student might hesitate before undertaking it. At the same
time, Richelieu’s personality and the times in which he lived are so
rich in varied interest that even a passing glance at both may be
found not unwelcome. If excuse is needed, there is that of Monsieur
de la Colombière: “Pour peu que j’en parle, j’en dirai beaucoup.”
There are many good authorities for the life of Cardinal de
Richelieu and for the details of his time, among which the well-known
and invaluable works of M. Avenel and of the Vicomte G. d’Avenel
should especially be mentioned. But any modern writer on the
subject must, first and foremost, acknowledge a deep obligation to
M. Hanotaux, concerning whose unfinished Histoire du Cardinal de
Richelieu, extending down to the year 1624, one can only express
the hope that its gifted author may some day find leisure and
inclination to complete it.
E. C. P.
 CONTENTS
List of Authorities Pages xiii, xiv
PART I
CHAPTER I
The birth of Armand Jean du Plessis de Pages 1-9
Richelieu—The position of his family—His
great-uncles—His grandfather and
grandmother—His father, François de
Richelieu, Grand Provost of Henry III.—His
mother and her family—His godfathers—The
death of his father
CHAPTER II
Friends and relations—The household at Pages 10-15
Richelieu—Country life in Poitou
CHAPTER III
The University of Paris—The College of Pages 16-25
Navarre—The Marquis du Chillou—A change
of prospect—A student of theology—The
Abbé de Richelieu at Rome—His
consecration
PART II
CHAPTER I
A Bishop at the Sorbonne—State of France Pages 26-37
under Henry IV.—Henry IV., his Queen and
his Court—The Nobles and Princes—The
unhealthiness of Paris—The Bishop’s
departure
CHAPTER II
Richelieu arrives at Luçon—His palace and Pages 38-47
household—His work in the diocese—His
friends and neighbours
CHAPTER III
“Instructions et Maximes”—The death of Pages 48-62
Henry IV.—The difficult road to favour—Père
Joseph and the Abbey of Fontevrault
CHAPTER IV
Waiting for an opportunity—Political unrest— Pages 63-71
The States-General of 1614—The Bishop of
Luçon speaks
CHAPTER V
Richelieu appointed Chaplain to Queen Anne Pages 72-87
—Discontent of the Parliament and the
Princes—The royal progress to the south—
Treaty of Loudun—Return to Paris—Marie de
Médicis and her favourites—The young King
and Queen—The Duc de Luynes—Richelieu
as negotiator and adviser—The death of
Madame de Richelieu
CHAPTER VI
A contemporary view of the state of France— Pages 88-100
Barbin, Mangot, and Richelieu—A new
rebellion—Richelieu as Foreign Secretary—
The Abbé de Marolles—Concini in danger—
The death of Concini—The fall of the Ministry
—Horrible scenes in Paris—Richelieu follows
the Queen-mother into exile
CHAPTER VII
Richelieu at Blois—He is ordered back to his Pages 101-115
diocese—He writes a book in defence of the
Faith—Marriage of Mademoiselle de
Richelieu—The Bishop exiled to Avignon—
Escape of the Queen-mother from Blois—
Richelieu is recalled to her service
CHAPTER VIII
The Treaty of Angoulême—The death of Pages 116-130
Henry de Richelieu—The meeting at
Couzières—The Queen-mother at Angers—
Richelieu’s influence for peace—The battle of
the Ponts-de-Cé—Intrigues of the Duc de
Luynes—Marriage of Richelieu’s niece—The
campaigns in Béarn and Languedoc—The
death of Luynes—The Bishop of Luçon
becomes a Cardinal
PART III
CHAPTER I
Cardinal de Richelieu—Personal descriptions Pages 131-142
—A patron of the arts—Court intrigues—
Fancan and the pamphlets—The fall of the
Ministers—Cardinal de Richelieu First
Minister of France
CHAPTER II
Richelieu’s aims—The English alliance—The Pages 143-157
affair of the Valtelline—The Huguenot revolt
—The marriage of Madame Henriette—The
Duke of Buckingham
CHAPTER III
Peace with Spain—The making of the army Pages 158-175
and navy—The question of Monsieur’s
marriage—The first great conspiracy—
Triumph of Richelieu and death of Chalais
CHAPTER IV
Two famous edicts—The tragedy of Pages 176-192
Bouteville and Des Chapelles—The death of
Madame and its consequences—War with
England—The siege of La Rochelle
CHAPTER V
The Duc de Nevers and the war of the Pages 193-206
Mantuan succession—The rebellion in
Languedoc—A new Italian campaign—
Richelieu as Commander-in-Chief
CHAPTER VI
Illness of Louis XIII.—“Le Grand Orage de la Pages 207-216
Cour.”—The “Day of Dupes”
CHAPTER VII
Flight from France of the Queen-mother and Pages 217-233
Monsieur—New honours for Cardinal de
Richelieu—The fall of the Marillac brothers—
The Duc de Montmorency and Monsieur’s
ride to Languedoc—Castelnaudary—The
death of Montmorency—Illness and recovery
of the Cardinal
CHAPTER VIII
The Cardinal and his palaces—The château Pages 234-248
and town of Richelieu—The Palais-Cardinal
—Richelieu’s household, daily life, and
friends—The Hôtel de Rambouillet—
Mademoiselle de Gournay—Boisrobert and
the first Academicians—Entertainments at
the Palais-Cardinal—Mirame
CHAPTER IX
Conquests in Lorraine—The return of Pages 249-262
Monsieur—The fate of Puylaurens—France
involved in the Thirty Years’ War—Last
adventures of the Duc de Rohan—Defeat,
invasion, and panic—The turn of the tide—
Narrow escape of the Cardinal—The flight of
the Princes
 CHAPTER X
Palace intrigues—Mademoiselle de Hautefort Pages 263-275
—Mademoiselle de la Fayette—The affair of
the Val-de-Grâce—The birth of the Dauphin
—The death of Père Joseph—Difficulties in
the Church
CHAPTER XI
Victories abroad—The death of the Comte de Pages 276-290
Soissons—Social triumphs—Marriage of the
Duc d’Enghien—The revolt against the taxes
—The conspiracy of Cinq-Mars—The
Cardinal’s dangerous illness—He makes his
will—The ruin of his enemies—His return to
Paris
CHAPTER XII
The Cardinal’s last days—Renewed illness— Pages 291-298
His death and funeral—His legacies—The
feeling in France—The Church of the
Sorbonne
INDEX Pages 299-306
 LIST OF ILLUSTRATIONS
FACING PAGE
Cardinal de Richelieu. Triple Portrait by Frontispiece
Philippe de Champagne (National Gallery)
Henry IV. From an engraving after the 26
picture by François Porbus
Cloister at Champigny 34
From a photo by A. Pascal, Thouars.
The Majority of Louis XIII. (Louis XIII. and 68
Marie de Médicis). From the picture by
Rubens in the Louvre
From a photo by Neurdein, Paris.
Cardinal de Richelieu. Portrait by Philippe 132
de Champagne
From a photo by A. Giraudon, Paris.
Gaston de France, Duc d’Orléans. From 162
a contemporary portrait
From a photo by Neurdein, Paris.
Louis XIII. From a contemporary portrait 188
From a photo by Neurdein, Paris.
The Château de Richelieu. From an old 234
print
The Town of Richelieu. From an old print 238
Anne of Austria. From a miniature in the 268
Victoria and Albert Museum
Porte de Châtellerault, Richelieu 280
 From a photo by Imprimerie Photo-
Mécanique, Paris.
Tomb of Cardinal de Richelieu, by 294
Girardon, in the Church of the Sorbonne
From a photo by Neurdein, Paris.
 CHIEF AUTHORITIES
CONTEMPORARY

Lettres, Instructions Diplomatiques et Papiers d’État du

Cardinal de Richelieu. Recueillis et publiés par M. Avenel.
Mémoires du Cardinal de Richelieu. Édition Petitot et
Monmerqué.
Mémoires du Cardinal de Richelieu. New Edition. With
Notes, etc. (Société de l’Histoire de France.) Not completed.
Mémoires sur la Régence de Marie de Médicis, par
Pontchartrain. Édition Petitot et Monmerqué.
Mémoires de Bassompierre. Édition Petitot et Monmerqué.
Journal de Pierre de l’Estoile. Édition Petitot et Monmerqué.
Mémoires du Marquis de Montglat. Édition Petitot et
Monmerqué.
Mémoires de Madame de Motteville. Édition Riaux.
L’Histoire du Cardinal-Duc de Richelieu. L. Aubery.
Testament Politique du Cardinal-Duc de Richelieu.
Journal de M. le Cardinal-Duc de Richelieu. 1630, 1631.
Portraits des Hommes Illustres François. M. de Vulson,
Sieur de la Colombière.
Le Véritable Père Joseph, Capucin. 1704.
Histoire du Roy Henry le Grand. Hardouin de Péréfixe.
Mémoire d’Armand du Plessis de Richelieu, Evêque de
Luçon, 1607 ou 1610. Édition Armand Baschet.
 Description de la Ville de Paris. Germain Brice.
Les Historiettes de Tallemant des Réaux.
Etc., etc.

MODERN

Histoire du Cardinal de Richelieu. G. Hanotaux.

Histoire de France. H. Martin. Vol. xi.
Histoire de France. Michelet. Vols. xiii. and xiv.
Vie Intime d’une Reine de France, Marie de Médicis. L.
Batiffol.
Le Roi Louis XIII. à Vingt Ans. L. Batiffol.
Louis XIII. et Richelieu. Marius Topin.
Richelieu et les Ministres de Louis XIII. B. Zeller.
La Noblesse Française sous Richelieu. Vicomte G.
d’Avenel.
Prêtres, Soldats et Juges sous Richelieu. Vicomte G.
d’Avenel.
Le Cardinal de Bérulle et le Cardinal de Richelieu. M.
l’Abbé M. Houssaye.
Gentilshommes Campagnards de l’Ancienne France. Pierre
de Vaissière.
Le Père Joseph et Richelieu. G. Fagniez.
Madame de Hautefort. Victor Cousin.
Madame de Chevreuse. Victor Cousin.
Le Règne de Richelieu. Émile Roca.
Le Cardinal de Richelieu: Étude Biographique. L. Dussieux.
Le Plaisant Abbé de Boisrobert. Émile Magne.
Etc., etc.
CARDINAL DE RICHELIEU