Pilot-scale production and purification of a staphylokinase-based fusion protein over-expressed in Escherichia coli

2008 ◽  
Vol 4 (1) ◽  
pp. 75-81 ◽  
Author(s):  
Genshen Zhong ◽  
Aiping Yu ◽  
Bingxing Shi ◽  
Yang Liu ◽  
Chutse Wu
KSBB Journal ◽  
2017 ◽  
Vol 32 (4) ◽  
pp. 300-305 ◽  
Author(s):  
Eun Jae Kim ◽  
Jun Hyuck Lee ◽  
Sung Gu Lee ◽  
Se Jong Han

2018 ◽  
Vol 81 (6) ◽  
pp. 953-962 ◽  
Author(s):  
HALEY S. SMOLINSKI ◽  
SIYI WANG ◽  
LIN REN ◽  
YUHUAN CHEN ◽  
BARBARA KOWALCYK ◽  
...  

ABSTRACT Several outbreaks of foodborne illness traced to leafy greens and culinary herbs have been hypothesized to involve cross-contamination during washing and processing. This study aimed to assess the redistribution of Salmonella Typhimurium LT2 during pilot-scale production of baby spinach and cilantro and redistribution of Escherichia coli O157:H7 during pilot-scale production of romaine lettuce. Four inoculated surrogate:uninoculated product weight ratios (10:100, 5:100, 1:100, and 0.5:100) and three inoculation levels (103, 101, and 10−1 CFU/g) were used for the three commodities. For each of three trials per condition, 5-kg batches containing uninoculated product and spot-inoculated surrogate products at each ratio and inoculation level were washed for 90 s in a 3.6-m-long flume tank through which 890 L of sanitizer-free, filtered tap water was circulated. After washing and removing the inoculated surrogate products, washed product (∼23, 225-g samples per trial) was analyzed for presence or absence of Salmonella Typhimurium or E. coli O157:H7 by using the GeneQuence Assay. For baby spinach, cilantro, and romaine lettuce, no significant differences (P > 0.05) in the percentage of positive samples were observed at the same inoculation level and inoculated:uninoculated weight ratio. For each pathogen product evaluated (triplicate trials), inoculation level had a significant impact on the percentage of positive samples after processing, with the percentage of positive samples decreasing, as the initial surrogate inoculation level decreased. The weight ratio of contaminated:noncontaminated product plays an important role: positive samples ranged from 0% to 11.6% ± 2.05% and from 68.1% ± 33.6% to 100% among the four ratios at inoculation of 10−1 and 101 CFU/g, respectively. To our knowledge, this study is the first to assess the redistribution of low levels of pathogens from incoming product to leafy greens during processing and should provide important data for microbial risk assessments and other types of food safety analyses related to fresh-cut leafy greens.


Polymers ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 290
Author(s):  
Chih-Yu Cheng ◽  
Chia-Huang Tsai ◽  
Pei-Jyun Liou ◽  
Chi-Hang Wang

For pilot-scale production of chito-oligosaccharides, it must be cost-effective to prepare designable recombinant chitosanase. Herein, an efficient method for preparing recombinant Bacillus chitosanase from Escherichia coli by elimination of undesirable substances as a precipitate is proposed. After an optimized culture with IPTG (Isopropyl β-d-1-thiogalactopyranoside) induction, the harvested cells were resuspended, disrupted by sonication, divided by selective precipitation, and stored using the same solution conditions. Several factors involved in these procedures, including ion types, ionic concentration, pH, and bacterial cell density, were examined. The optimal conditions were inferred to be pH = 4.5, 300 mM sodium dihydrogen phosphate, and cell density below 1011 cells/mL. Finally, recombinant chitosanase was purified to >70% homogeneity with an activity recovery and enzyme yield of 90% and 106 mg/L, respectively. When 10 L of 5% chitosan was hydrolyzed with 2500 units of chitosanase at ambient temperature for 72 h, hydrolyzed products having molar masses of 833 ± 222 g/mol with multiple degrees of polymerization (chito-dimer to tetramer) were obtained. This work provided an economical and eco-friendly preparation of recombinant chitosanase to scale up the hydrolysis of chitosan towards tailored oligosaccharides in the near future.


2015 ◽  
Vol 173 ◽  
pp. 827-837 ◽  
Author(s):  
Domien De Paepe ◽  
Katleen Coudijzer ◽  
Bart Noten ◽  
Dirk Valkenborg ◽  
Kelly Servaes ◽  
...  

2017 ◽  
Vol 109 ◽  
pp. 634-645 ◽  
Author(s):  
Svitlana Nitièma-Yefanova ◽  
Valérie Tschamber ◽  
Romain Richard ◽  
Sophie Thiebaud-Roux ◽  
Brice Bouyssiere ◽  
...  

2008 ◽  
Vol 13 (2) ◽  
pp. 182-188 ◽  
Author(s):  
Kang-Ik Jo ◽  
You-Jung Lee ◽  
Bo-Kyung Kim ◽  
Bo-Hwa Lee ◽  
Chung-Han Chung ◽  
...  

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