Shung-Chang Jong's research while affiliated with ATCC and other places

Introduction Historical Outline Immunomodulatory and Antitumor Glycans β‐Glucans and their Protein Complexes Heteroglucans and their Protein Complexes Chemical Modification Antiviral Glycans Antimicrobial Glycans Hepatoprotective Glycans Antifibrotic Glycans Antiinflammatory Glycans Antidiabetic and Hypoglycemic Glycans Hypocholesterolemic Glycans Patents Outlook and Perspectives

ITS and partial β-tubulin genes of 17 ATCC reference strains of Monascus species were PCR amplified and sequenced. Monascus pilosus and M. ruber could not be differentiated with these sequences, suggesting a synonymy. In maximum parsimony analyses on both data sets, M. ruber, M. pilosus, M. purpureus, and M. sanguineus were placed into the same clade. ITS sequence alignment revealed a number of gaps in ITS1 and ITS2 of M. pallens, M. lunisporas, and M. eremophilus compared to M. purpureus, M. ruber, and M. pilosus. Accordingly, analyses with the ITS sequences placed these species into clades, incongruent with the analyses using the partial β-tubulin genes and the previous results with the partial large subunit rRNA genes. The phylogenetic relationship derived from the partial β-tubulin genes was similar to those postulated by the 5'-partial LSU rRNA genes. This finding strongly suggests that evolutionary or phylogenetic classification with ITS sequence information should be performed with caution. In the phylogenetic trees with the ITS sequences, M. lunisporas was distantly associated with Aspergillus ustus; M. pallens was placed in a clade that shares a common node with A. versicolor; and M. eremophilus was placed on a branch separate from the M. purpureus, M. ruber, and M. pilosus group while M. pallens and M. lunisporas were placed into the related clades sharing a common node in the tree derived from the partial β-tubulin gene. Each of the phylogenetic analyses with the partial β-tubulin genes, the ITS, or the 5'-end of the LSU rRNA, as previously carried out, placed M. eremophilus into a different lineage. Molecular analyses with these molecular targets generated three different topologies for M. eremophilus, indicating a unique and unpredictable genetic combination for this species. It might reflect extreme environmental stress on this species and subsequent genetic changes.

ITSs and D1/D2 regions of the LSU rRNA genes of 57 ATCC strains of Cladosporium representing common airborne species, C. herbarum, C. cladosporioides and C. sphaerospermum, were PCR-amplified and sequenced in both directions. Sequence alignments and subsequent maximum parsimony analyses on both of the datasets indicated that C. herbarum and C. cladosporioides were more closely related to each other than either was to C. sphaerospermum. Most strains were clustered into one of the three individual groups according to the species identification by the depositors. However, four strains deposited as C. herbarum clustered with C. clasdosporioides. Conidial shapes of these four strains resembled C. herbarum although their size was smaller. Another strain deposited as C. cladosporioides clustered with C. herbarum. The conidia of this strain were longer than the typical conidia of C. cladosporioides, but resembled C. cladosporioides. ATCC 64726 (deposited as C. cladosporioides) and ATCC 26362 (deposited as C. herbarum) were placed close to C. sphaerospermum, but formed a distinct subgroup with strong bootstrap supports. The strains of these three Cladosporium species collected over a wide range of areas displayed minute intra-species sequence variations and varying degrees of conidial shapes and sizes.

 The D1/D2 regions of the large subunit (LSU) rRNA genes of 65 strains of Monascus and Xeromyces were PCR amplified and sequenced in both directions. Maximum-parsimony analysis produced five most parsimonious trees. The strict consensus tree of these five parsimonious trees clustered M. eremophilus, M. ruber, M. pilosus, M. purpureus, and M. sanguineus in the same clade, reflecting high sequence similarity. M. sanguineus, M. purpureus, M. ruber, and M. pilosus differed in one or two nucleotides. The sequence of M. eremophilus ATCC 62925 isolated from a xerophilic environment differed from M. purpureus in only one nucleotide, despite pronounced morphological and ecological differences when compared with the other species. M. lunisporas, M. floridanus, M. pallens, and X. bisporus were each placed in a separate branch, confirming their taxonomic descriptions as individual species. Maximum-likelihood analysis on the same data set generated a single tree and grouped the species of the first clade in the parsimony analysis into a single clade but placed the rest of the Monascus species and Xeromyces bisporus on different branches. The trees inferred from both analyses revealed a monophyletic relationship between Monascus and Xeromyces, when compared with other related cleistothecial or imperfect genera.

A crude polysaccharide-protein complex from Lactarius deliciosus fruit bodies (LDP) was extracted with hot water, precipitated, and washed with 95% ethanol. After protein removal according to the Sevag method, LDP was dialyzed against running tap water and distilled water, and further purified by Sephadex G-100 column chromatography. Gas chromatography showed that LDP is composed of glucose, galactose, mannose, and arabinose in a molar ratio of 100:14.9:6.23:1.97. According to Gel Permeation Chromatography-HPLC results, the molecular weight is 642kDa. An IR spectrum revealed coinciding polysaccharide and peptide absorption peaks, while a 1H-NMR spectrum indicated the main chain has a ß-pyranglycoside linkage. Amino acid analysis detected the presence of 17 kinds of amino acids.

Pure culture methodology allows an investigator to detect, isolate, identify, and quantify numbers and kinds of fungi from a wide array of environments and to define the nutritional, chemical, and environmental requirements for their growth and metabolism. Studies with pure cultures not only enhance understanding of any natural ecosystem within infected hosts or in the environment, but also help in the determination of molecular architecture and screening for novel activity/compounds. Pure culture methodology is the foundation of fundamental and applied research, as well as the commercial exploitation of fungi.

The yeast Schwanniomyces occidentalis produces a killer toxin lethal to sensitive strains of Saccharomyces cerevisiae. Killer activity is lost after pepsin and papain treatment, suggesting that the toxin is a protein. We purified the killer protein and found that it was composed of two subunits with molecular masses of approximately 7.4 and 4.9 kDa, respectively, but was not detectable with periodic acid-Schiff staining. A BLAST search revealed that residues 3 to 14 of the 4.9-kDa subunit had 75% identity and 83% similarity with killer toxin K2 from S. cerevisiaeat positions 271 to 283. Maximum killer activity was between pH 4.2 and 4.8. The protein was stable between pH 2.0 and 5.0 and inactivated at temperatures above 40°C. The killer protein was chromosomally encoded. Mannan, but not β-glucan or laminarin, prevented sensitive yeast cells from being killed by the killer protein, suggesting that mannan may bind to the killer protein. Identification and characterization of a killer strain of S. occidentalis may help reduce the risk of contamination by undesirable yeast strains during commercial fermentations.

Any living biological material is, in effect, a biotechnology factory complete with sophisticated process designs and its own source of venture capital. Biotechnology is the application of living material and its process system to obtain useful products or services. It is not limited to the recent developments in recombinant DNA techniques, monoclonal antibodies, and related processes and products, aspects with which it is most often identified, but includes the manufacture of nonrecombinant biological material, such as microorganisms and plants and animals from traditional breeding methods; biologically active compounds; and a variety of foods and drugs. Due to complex economic, social, and technological changes, biotechnology is in a state of transition from a knowledge-building science to a new industry. Among the many issues and concerns contributing to this is the emergence of ownership and intellectual property rights of biological material. Since the biotechnology industry is becoming increasingly global in nature, competition in world markets requires that special attention be paid to regulations concerning deposit requirements for patent purposes, transfer agreements, and the packing and shipping of biological material, both nationally and internationally.

Restriction fragment length polymorphisms (RFLPs) in two regions of the ribosomal DNA (rDNA) repeat unit were examined in 33 strains representing 18 species of Saprolegnia. The Polymerase Chain Reaction (PCR) was used to separately amplify the 18S rDNA and the region spanning the two internal transcribed spacers (ITS) and the 5.8S ribosomal RNA gene. Amplified products were subjected to a battery of restriction endonucleases to generate various fingerprints. The internal transcribed spacer region exhibited more variability than the 18S rDNA and yielded distinctive profiles for most of the species examined. Most of the species showing 100% similarity for the 18S rDNA could be distinguished by 5.8S + ITS restriction polymorphisms except for S. hypogyna, S. delica, S. lapponica, and S. mixta. The rDNA data indicate that S. lapponica and S. lapponica and S. mixta are conspecific with S. ferax, whereas there is no support for the proposed synonymies of S. diclina with S. delica and of S. mixta with S. monoica. Results from cluster analysis of the two data sets were very consistent and tree topologies were the same, regardless of the clustering method used. A further examination of multiple strains in the S. diclina-S. parasitica complex showed that restriction profiles are conserved across different strains of S. parasitica originating from the U.K. and Japan. HhaI and BsaI restriction polymorphisms were observed in isolates from the U.S. and India. The endonuclease BstUI was diagnostic for S. parasitica, generating identical fingerprints for all stains regardless of host and geographic origin. Except for the atypical strain ATCC 36144, restriction patterns were also largely conserved in S. diclina. Correlation of the rDNA data with morphological and ultrastructural features showed that S. diclina and S. parasitica are not conspecific. Restriction polymorphisms in PCR-amplified rDNA provide a molecular basis for the classification of Saprolegnia and will be useful for the identification of strains that fail to produce antheridia and oogonia.