60 BIOTECHNOLOGY AND BIOENGINEERING

References 1. Champignat, A., C. Vernet, B. Laine, and J. Filosa, Nature, 197, 13 (1963). 2. Filosa, J. (to British Petroleum Company, Ltd.), British Patent 914,568. 3. Hedrick, H. G., M. T. Carroll, H. P. C. Owen, and D. J. Pritchard, Appl.

4. Hitzmann, D. O., and A. hl. Mills (to Phillips Petroleum Co.), U.S. Patent

5. Hosler, P., Biotechnol. Bioag., 5 , 243 (1963). 6. Kersten, D. Ch., Mikrobiologiya, 32, 1024 (1963). 7. Kester, A. S., and J. W. Foster, J. Bacteriol., 85,859 (1963). 8. Lukins, H. B., and J. W. Foster, 2. Allgem. Mikrobiol., 3,251 (1963). 9. Miller, T. L., and M. J. Johnson, in Abstracts, 145th Meeting, Am. Chem.

10. Molson, R. K., J. N. Baptist, and M. J. Coon, Biochemistry, 2,1155 (1963). 11. Payne, W. J., and V. E. Feisal, Appl. Microbiol., 11, 339 (1963). 12. Phillips, U. A., and R. W. Traxler, Appl. Microbiol., 11, 235 (1963). 13. Shiio, I. S., Otsuka, R. Ishii, N. Katsuya, and H. Iizuka, J . Gen. Appl.

14. Slavnina, G. P., Mikrobiologiya, 32, 121 (1963). 15. SociCt6 Frangaise des PCtroles B. P., French Patent 1,297,619, May 21,

16. SociCtC Frangaise den PCtroles B. P., French Patent 1,320,058, Jan. 28, 1963. 17. Stone, R. W., Develop. Ind. Microbiol., 4.5 (1963). 18. Sturm, L. C., and E. P. Rosanova, Mikrobiologiya, 32,1013 (1963). 19. Takahashi, J., K. Kobayashi, Y. Kawabata, and K. Yamada, Agr. Biol.

20. Thijsse, G. J. E., and A. C. van der Linden, Anlonie van Leeuwenhoek, 27,

21. Yamada, K., K. Kobayashi, J. Takahashi, and Y. Imada, Agr. Biol. Chem.

22. Yamada, K., J. Takahashi, and K. Kobayashi, Nature, 198,1115 (1963). 23. Yamada, K., J. Takahashi, and K. Kobayashi, Agr. Biol. Chem. (Tokyo),

Microbiol., 11, 472 (1963).

3,084,106, April 2, 1963.

Soe., New York, 1964.

Microbiol. (Tokyo), 9.23 (1963).

1962.

Chem. (Tokyo), 27,839 (1963).

89 (1963).

(Tokyo), 27, 390 (1963).

27,773 (1963).

Miscellaneous Products

Alkaloids

A patent was issued for the production of lysergic amide deriva- tives by Claviceps pa~pali .~R Another patent was issued for the preparation of lysergic acid or its derivatives by growth of Aspergillus clavatus on a medium containing 5% mannitol, 0.54% succinic acid,

BIOTECHNOLOGY A N D BIOENGINEERING, VOL. VII. ISSUE 1

FERMENTATION REVIEW 61

and ~ a l t s . 4 ~ The biosynthesis of the ergot alkaloids was the subject of several papers issuing from the Prairie Regional Laboratory. It was shown that alkaloid production depends on continued respiration but noncontinued accumulation of shunt products during the late growt,h phase. Supplementation with glucose resulted in decreased alkaloid accumulatior1.~-5~ The incorporation of m-tryptophane- 14C, but not generally labelled L-phenylalanine or L-tyrosine, into the ergoline moiety of ergot alkaloids was demonstrated. Small amounts of me~alonate-2-~~C or formate-I4C could aIso be in~orpora ted .~~

The selection of monohyphal isolates of Claviceps paspali according to their ability to produce lysergic acid alkaloids resulted in a strain which could accumulate up to 645 mg./l. total lysergic acid alkaloids as compared to 50 mg./l. for the original strain. The addition of 0.5 g./l. of tryptophan or 10 g./L acetaiiiide increased the figure by a fac- tor of about Another paper reported that the addition of 3% 1,Zpropanediol to the medium increased the yield of alkaloids from about 390 mg./I. to 415 n1g./l.13

The transformation of various yohimbine alkaloids was described. Calonectria decora, Cunninghamella echinulata, Cu. bertholleliae, and Streptomyces rimosus were capable of hydroxylating yohimbine, a- yohimbine, and @-yohimbine in the 10, ll, and 18 position^.^^

Patents were issued for the preparation of demethylated deriva- tives of colchicine by cultivation of species of Streptomyces, Fusarium, Curvularia, Corynebacterium, Cunninghamella, Aspergillus, and Strep- tococcus followed by addition of the colchicine compound to be de- niethylated. r 41

Gibberellins Several patents were issued for the production of gibberellins in

submerged culture by Gibberella fujikuroi. A typical niedium con- sisted of glucose, 15%; ammonium nitrate, 0.25% ; and salts.10~21-23 Another patent was issued for a process in which the medium con- tained 4-12% dextrin and 2-1075 sucrose totalling together 1&15%.l An unusual process consisted of growing G. fujikuroi in a two phase system of crude oil and water. Acids, aldehydes, alcohols, and ke- tones are produced in the organic phase and gibberellins in the aqueous phase. l5

Using Fusarium monilzforme, yields of up to 1.2 g./l. of gibberellic acid were produced on a medium composed of glucose, 20; corn steep liquor, 25; ammonium nitrate, 2.6 g./l.; and salts.42

62 BIOTECHNOLOGY AND BIOENGINEERING

The presence of gibberellin-like substances in various species of algae was reported.32

Insecticides Several patents were issued to Bioferm C ~ r p . ’ , * * ~ ~ They claimed

the production of Bacillus thuringiensis in a medium formulated such that the assimilable carbohydrate and nitrogen are exhausted within 6 hr. of the start of sporulation. Population sporulations of up to 95% were obtained. A typical medium contained 1.4% beet mo- lasses, 1.2% corn steep solids, and 0.1% calcium carbonate. An- other patent described the production of B. thuringiensis by first ab- sorbing an inoculuni on a suitable material, passing humidified air a t 30°C. over it for 2 4 8 hr. until sporulation occurs, and then iri- creasing the air temperature so as to dry the material below a mois- ture content of 1070.29 Still another patent described the produc- tion of B. thuringiensis on a medium containing 1-2% glucose and 0.064.1~o assimilable nitrogen.24 A series of patents described the use of Pseudonzonas desmolytica, P. dacunhae, Aerobacter strains, and Achromobacter strains to destroy nematodes.16-20

A factor in B. thuringiensis different from the crystalline inclusion and toxic to flies was described as having some promise as a feed additive for livestock.30 Another report described the effect of this microorganism on honey bee population^.^^ Two reports were con- cerned with B. popilliae. In one, a medium suitable for sporulation of this organism was described. It required a large amount of certain types of yeast extract, the absence of glucose and the presence of ace- tate, and a narrow pH Both the hexosemonophosphate and Emden-Yleyerhof pathways were found, the participation of the first being about 1-4Oy0 dependirig on the amount of oxygen in the at- mosphere in the re~pi rometer .~~ An unidentified growth factor from agar was shown to permit growth of B. lentimorbus on a tryptone- glucose-yeast extract medium which itself could not support growth. l4

Other Products

A process for converting part of the glucose in invert syrup to glu- conic acid by means of Acetobacter suboxydans was described. The fructose was ~ n a t t a c k e d . ~ ~ Another report indicated that a medium poor in vitamins was preferable for the conversion of sorbitol to sor-

BIOTECHNOLOGY AND BIOENGINEERING. VOL. vn. ISSUE I

FERMENTATION REVIEW 63

bose by A . suboxydans, as it increased the oxidation with respect to cell proliferation.55 A phosphat,e concentration of 10-50 mg./l. was also found optimal.45

A patent was issued for the reduction of ket.ocarboxylic acids to optically active lactones and corresponding hydroxy carboxylic acids. The medium contained fermentable substrate and the keto acids were added gradually. Various yeasts, molds, or bacteria could be

Several reports were concerned with microbiological leaching of ores, a process of potential significance in the mining industry. Var- ious thiobacilli and ferrobacilli are able to oxidize sulfide to sulfate, produce acid, oxidize ferrous to ferric, and thus solubilize copper resi- dues in waste mineral heaps. The soluble copper can be recovered by precipitation. 11,38,39

An extremely toxic metabolite produced by Aspergillus jlavus was identified.2 It could also be produced by the closely relat,ed A . p a r a s i t i c ~ s . ~ Streptomyces venexuelae produced D-t.hreo-l-p-aniino- pheny1-2-dichloroacetamide-1,S-propanediol in addition to chlor- aniphenicol under normal fermentation condition^.^^

Other reports dealt with the conversion of benzaldehyde to phenyl- a~etylcarbinol ,~.~ the oxidat,ion of glycerine to dihydroxyacetone by Actinomyces aureofa~iens,3~ produc.tior1 of a green iron containing pig- ment, ferroverdin, by a Streptomyces ~pecies ,~ the preparation of radioactrive biochemicals from yeast,28 degradation of quercet.inS4 and rutin,(? biosynthesis of d i a~e ty l ,~* and lysing microorganisms with hypertonic strontium salts, l2 enzynles and antibiotics and surface active agents,27 or treatment with N-glucanase to recover cell coin- ponents.26

References 1. Abbott Laboratories, British Patent 919,186, Feb. 20, 1963. 2. Asao, T., G. Buchi, 11. 1%. Abdel-Kader, S. B. Chang, E. L. Wick, and

3. Ballio, A., H. Bertholdt, A. Carilli, E. B. Chain, V. DiVittorio, A. Tonolo,

4. Bellet, P., and L. Vellus (to Roussel-UCLAF), U.S. Patent 3,090,729, May

5. BeCvBFovB, H., and 0. HanE, Foliu Microbiol. (Prague), 8,42 (1963). 6. BeEvMovB, H., 0. H a d , and K. Macek, Folia Microbiol. (Prague), 8,

G. N. Wogan, J . Am. Chem. Soc., 85, 1706 (19631.

and L. Vero-Barcellona, Proc. Royal SOC. (London), Ser. B, 158,43 (1963).

21, 1963.

165 (1963).

64 BIOTECHNOLOGY AND BIOENGINEERING

7. Bioferm Corp., German Patent 1,158,746, Dec. 5, 1963. 8. Bioferm Corp., Japanese Patent 9946, June 21, 1963, 9. Codner, R. C., K. Sergeant, and R. Yeo, Biotechnol. Bioeng., 5 , 185 (1963).

10. Cross, B. E., and J. R. Hanson (to Imperial Chem. Industries, Ltd.),

11. Ehrlich, H. L., J. Bacteriol., 86, 350 (1963). 12. Fujisawa Pharmaceutical Co., Ltd., Japanese Patent 9934, June 21, 1963. 13. Groger, D., and V E. Tyler, Jr., Lloydia, 26, 174 (1963). 14. Haynes, W. C., and L. Rhodes, Bacteriol. Proc., 1963, 10. 15. Hitzmann, D. O., and A. 31. Mills (to Phillips Petroleum Co.), U.S.

16. Ihara Agricultural Chemicals, Ltd., Japanese Patent 3687, April 18, 1963. 17. Ihara Agricultural Chemicals, Ltd., Japanese Patent 3688, April 18, 1963. 18. Ihara Agricultural Chemicals, Ltd., Japanese Patent 3689, April 18, 1963. 19. Ihara Agricultural Chemicals, Ltd., Japanese Patent 3690, April 18, 1963. 20. Ihara Agricultural Chemicals, Ltd., Japanese Patent 3691, April 18, 1963. 21. Imperial Chemical Industries, Ltd., German Patent 1,150,347, June 20,

22. Imperial Chemical Industries, Ltd., German Patent 1,150,348, June 20,

23. Imperial Chemical Industries, Ltd., German Patent 1,155,413, Oct. 10,

24. Institiit Pasteur, British Patent 942,462, Nov. 20, 1963. 25. Kett, G. (to J. and S. Colman, Ltd.), Britivh Patent 923,858, 1963. 26. Kinki Yakult Seizo, K. K., and Meiji Seika Kaisha, Ltd., French Patent

27. Kyowa Hakko Kogyo Co., Ltd., Japanese Patent 9931, June 21, 1963. 28. Laufer, L., S. Gutcho, T. Castro, and R. Grennen, in Abstracts, 145th Meet-

29. Mechalas, B. J. (to Nutrilite Products, Inc.), U.S. Patent 3,086,922, April

30. Mechalas, B. J., and 0. Beyer, DeveZop. Ind. Microbiol., 4, 142 (1963). 31. Megna, J. (to Bioferm Corp.), U.S. Patent 3,073,749, Jan. 15, 1963. 32. Mowat, J. A., Nature, 200,453 (1963). 33. Muys, G. T., B. van der Ven, and A. P. Jonge (to Unilever, Ltd.), British

34. Pacifici, L. R., W. J. Keileher, and A. E. Schwarting, Lloyd&, 26. 161

35. Patterson, E. L., W. W. Andres, E. F. Krause, R. E. Hartman, and L. A.

36. Pepper, R. E., and R. N. Costilow, Bacteriol. Proc., 1963, 10. 37. Plakunova, V. G., Dokl. Akad. Nauk. SSSR, 148,445 (1963). 38. Razzell, W. E., and P. C. Trussell, Appl. Microbiol., 11, 105 (1963). 39. Razzell, W. E., and P. C. Trussell, J. Bacteriol., 85, 595 (1963). 40. Rhodes, R. A,, M. Shekleton, G. R. Hrubant, H. H. Hall, and R. W. Jack-

41. Roussel-UCLAF, British Patent 923,421, April 10, 1963.

British Patent 914,893, Jan. 9, 1963.

Patent 3,084,106, April 2, 1963.

1963.

1963.

1963.

1,333,739, Aug. 2, 1963.

ing, Am. Chem. Soc., New York, Sept. 9, 1963.

24, 1963.

Patent 916,822, 1963.

(1963).

Mitscher, Arch. Biochem. Biophys., 103, 117 (1963).

son, Bucteriol. Proc., 1963, 10.

BIOTECHNOLOGY AND BIOENGINEERING, VOL. VII, ISSUE 1

FERMENTATION REVIEW 65

42. Sanchez-Marroquin, A., A p p l . Microbwl., 11, 523 (1963). 43. Sandoz S. A., Belgian Patent 629,158, Oct. 21, 1963. 44. Seitz, S. W., W. E. Sandine, P. R. Elleker, and E. A. Day, Can. J . Micro-

45. Shchekunova, S. A., Mikrobiologiya, 32, 529 (1963). 46. Shimanuki, H., and P. A. Hartman, Bacteriol. Proc., 1963, 10. 47. Simpson, F. J., N. Narasimhachari, and D. W. S. Westlake, Can. J . Mi-

48. Societh Farmaceutici Italia, German Patent 1,140,670, Dec. 6, 1963. 49. Stratton, C. D., and M. C. Rebstock, Arch. Bwchem. Bwphys., 103, 159

50. Taber, W. A., Bacteriol. Proc., 1963, 23. 51. Taber, W. A., Develop. Ind. Microbiol., 4, 295 (1963). 52. Taber, W. A., and L. C. Vining, Can. J . Microbiol., 9 , 1 (1963). 53. Vining, L. C., and W. A. Taber, Can. J . Microbiol., 9, 291 (1963). 54. Westlake, D. W. S., Can. J. Microbwl., 9, 211 (1963). 55. Zhdan-Pushkina, S. M., and R. A. Kreneva, Mikrobiologiya, 32,711 (1963).

biol., 9, 431 (1963).

crobiol., 9, 15 (1963).

(1963).

Nucleosides and Nucleotides

5’-Nucleotides

Methods for producing these chemicals underwent extensive in- vestigation as indicated by the increase in reports during 1963. Certain 5’-nucleotides, notably inosinic, guanylic, and xanthylic acids appear to have some promise as food flavor extenders, particu- larly for beef-like tastes. While they have attained some com- mercial success in Japan, their reception in other markets has been hindered by their high price.

A number of microbial routes for their preparation is available. They include : (1 ) the 5’-phosphodiesterase hydrolysis of ribonucleic acid usually of microbial origin ; (2) direct fermentation biosynthe- sis; (3) enzymatic phosphorylation of microbially produced ribo- nucleosides; and (4) deamination of adenylic acid, a nonflavor ex- tending 5‘-nucleotide produced by one of the previously enumerated routes.

A 300 reference review dealing with the production of nucleic acids and their derivatives was omp piled.^? Methods for the selection of microbes producing these materials were described, from among 925 different strains, froin 75 genera and 370 species,34 from among 407