title: Dietary supplementation with bovine lactoferrampin-lactoferricin produced by Pichia pastoris fed-batch fermentation affects intestinal microflora in weaned piglets.
abstract: This work is aimed at investigating the effects of recombinant bovine lactoferrampin-lactoferricin (LFA-LFC) instead of chlortetracycline on intestinal microflora in weaned piglets. The high cost of peptide production from either native digestion or chemical synthesis limits the clinical application of antimicrobial peptides. The expression of recombinant peptides in yeast may be an effective alternative. In the current study, recombinant LFA-LFC was produced via fed-batch fermentation in recombinant strain Pichia pastoris (KM71) XS10. Uniform design U6(6(4)) was used to optimize the fermentation conditions. The target peptide purified via cation-exchange and size-exclusion chromatography was added into the dietary of weaned piglets. After 21 days, the Lactobacilli, Bifidobacteria, and Enterobacteria in the chyme of the gut were quantified using real-time polymerase chain reaction. The results showed that approximately 82 mg of LFA-LFC was secreted into 1 L of medium under optimized conditions. Moreover, purified peptide showed strong antimicrobial activities against all the tested microorganisms. Compared with the control group, the LFA-LFC group increased the amount of Lactobacilli and Bifidobacteria (P<0.05) in the chyme of the stomach, duodenum, jejunum, ileum, colon, and caecum. These results show that dietary supplementation with LFA-LFC can affect intestinal microflora in weaned piglets.
authors.affiliation: Scientific Observing and Experimental Station of Animal Nutrition and Feed Science in South-Central, Ministry of Agriculture, Hunan Provincial Engineering Research Center of Healthy Livestock, Key Laboratory of Agro-ecological Processes in Subtropical Region, Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha, Hunan 410125, China.
authors.forename: Xiang-Shan
authors.initials: XS
authors.lastname: Tang
authors.forename: Hua
authors.initials: H
authors.lastname: Shao
authors.forename: Tie-Jun
authors.initials: TJ
authors.lastname: Li
authors.forename: Zhi-Ru
authors.initials: ZR
authors.lastname: Tang
authors.forename: Rui-Ling
authors.initials: RL
authors.lastname: Huang
authors.forename: Sheng-Ping
authors.initials: SP
authors.lastname: Wang
authors.forename: Xiang-Feng
authors.initials: XF
authors.lastname: Kong
authors.forename: Xin
authors.initials: X
authors.lastname: Wu
authors.forename: Yu-Long
authors.initials: YL
authors.lastname: Yin
mesh_terms: D000818:Animals; D002417:Cattle; D019587:Dietary Supplements; D054307:Enterotoxigenic Escherichia coli / Q000187:drug effects / Q000502:physiology; D005285:Fermentation; D007422:Intestines / Q000382:microbiology*; D007781:Lactoferrin / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D010446:Peptide Fragments / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D010843:Pichia / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D013552:Swine; D014886:Weaning
chemical_list: D010446:Peptide Fragments; C502040:lactoferrampin; C082171:lactoferricin B; D007781:Lactoferrin