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Title: Expression and secretion of active Moringa oleifera coagulant protein in Bacillus subtilis
Authors: Arnett, Clint M.
Lange, Justin.
Boyd, Ashley N.
Page, Martin A.
Cropek, Donald M. (Donald Michael)
Keywords: Bacillus subtilis
Coagulant protein
Moringa oleifera
Water treatment
Water turbidity
Publisher: Construction Engineering Research Laboratory (U.S.)
Engineer Research and Development Center (U.S.)
Series/Report no.: Miscellaneous Paper (Engineer Research and Development Center (U.S.)) ; no. ERDC/CERL MP-21-2
Is Version Of: Arnett, Clint, Justin Lange, Ashley Boyd, Martin Page, and Donald Cropek. "Expression and secretion of active Moringa oleifera coagulant protein in Bacillus subtilis." Applied Microbiology and Biotechnology 103, no. 23 (2019): 9411-9422.
Abstract: Cationic polypeptide proteins found in the seeds of the tropical plant Moringa oleifera have coagulation efficiencies similar to aluminum and ferric sulfates without their recalcitrant nature. Although these proteins possess great potential to augment or replace traditional coagulants in water treatment, harvesting active protein from seeds is laborious and not cost-effective. Here, we describe an alternative method to express and secrete active M. oleifera coagulant protein (MO) in Bacillus subtilis. A plasmid library containing the MO gene and 173 different types of secretory signal peptides was created and cloned into B. subtilis strain RIK1285. Fourteen of 440 clones screened were capable of secreting MO with yields ranging from 55 to 122 mg/L of growth medium. The coagulant activity of the highest MO secreting clone was evaluated when grown on Luria broth, and cell-free medium from the culture was shown to reduce turbidity in a buffered kaolin suspension by approximately 90% compared with controls without the MO gene. The clone was also capable of secreting active MO when grown on a defined synthetic wastewater supplemented with 0.5% tryptone. Cell-free medium from the strain harboring the MO gene demonstrated more than a 2-fold reduction in turbidity compared with controls. Additionally, no significant amount of MO was observed without the addition of the synthetic wastewater, suggesting that it served as a source of nutrients for the effective expression and translocation of MO into the medium.
Description: Miscellaneous Paper
Gov't Doc #: ERDC/CERL MP-21-2
Rights: Approved for Public Release; Distribution is Unlimited
Size: 18 pages / 687.03 kB
Types of Materials: PDF/A
Appears in Collections:Miscellaneous Paper

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