Bacto tryptone and yeast extract

Yeast nitrogen base (YNB) was obtained from Sunrise Biosciences, and Bacto™ Tryptone and Yeast Extract was purchased from Becton Dickinson and Company (Franklin Lakes, NJ). Antifoam 204 was obtained from Sigma-Aldrich (St. Louis, MO). Certified ACS grade crystalline sucrose was purchased from Fisher Scientific (Pittsburg, PA). Boc-triglycine was supplied by Bachem Americas, Inc. (Torrance, CA). The enzymes PNGase F, Sortase, and B. thetaiotaomicron α-1,2-mannosidase (BT3990, B.t. α-1,2-mannosidase) were produced inhouse.^{52 (link)–55 (link)} Endoglycosidase H and restriction enzymes were obtained from New England Biolabs (Ipswich, MA). Protein G resin was produced by coupling protein G (recombinantly expressed in E. coli)^{56 (link)} with Sepharose^® CL-4B (Sigma-Aldrich, St Louis, MO) using divinyl sulfone as a coupling reagent.⁵⁷ General chemicals were purchased from Sigma-Aldrich and Fisher Scientific unless otherwise noted.

Adenosine-3′-phosphate-5′-phosphosulfate (PAPS), adenosine-3′,5′-diphosphate sodium salt (PAP), potassium phosphate monobasic, phenylmethylsulfonyl fluoride (PMSF), and dehydroepiandrosterone (DHEA) were purchased from Sigma-Aldrich (St. Louis, MO). PAPS was further purified (>98% as analyzed by HPLC) using a previously described procedure (Sekura 1981 ). Bacto tryptone and yeast extract were obtained from Becton Dickinson, Co. (Sparks, MD) and Hydroxyapatite (Bio-Gel HT) from Bio-Rad Laboratories (Hercules, CA). Tween 20 was purchased from J.T. Baker Chemicals (Phillipsburg, NJ). Isopropyl-1-thio-D-galactopyranoside (IPTG) was from AMRESCO Inc. (Solon, OH). All other chemicals used were of the highest purity commercially available.