Lecture 1

Question 1

Define biotechnology

Answer 1

Application of scientific and engineering principles to the processing of materials by biological agents to provide goods and services

Question 2

What is fermentation?

Answer 2

The chemical transformation of organic components with aid of enzymes, especially microbial

Question 3

Five types of antique fermentation still around

Answer 3

Wine production – religious texts; Spirit production – 800BC china distillation of sake, EU distillation by 11/12th century, d. fermented grain global 14th cent. AD; Breadmaking – 6000BC Sumerians Babylonians make beer, 4000BC Egyptions brewers least leaven bread; Vinegar and yoghurt – acetic acid and lactic acid fermentation; Cheese and fermented milk products – bacterial and fungal mould

Question 4

When was the existence of microbes first recognised and by whom?

Answer 4

In 17th cen. 1676 by dutch microscopist Antonie van Leeuwenhoesk, described ‘animalcules’ = bacteria in microscope , sig as fermentation requires microbes

Question 5

Who showed that microbes could NOT arise spontaneously from nothing and when?

Answer 5

Louis PASTEUR and John TYNDALL in the 1860s

Question 6

What else did PASTEUR discover?

Answer 6

Each type of fermentation mediated by specific microbe, possible for microbes to be anaerobic

Question 7

What scientists were primarily involved in the start of biotechnology as a science?

Answer 7

Antonie van Leeuwenhoesk 1676 animalcules; PASTEUR and TYNDALL 1860s non spontaneous microbes; KOCH grew bacteria in lab pure culture; PASTEUR fermentation by specific microbe, strictly anaerobic; BUCHNER 1897 cell-free metabolism -> enzymes

Question 8

How and who discovered enzymes?

Answer 8

Edward BUCHNER 1897 –filtered extract of macerated yeast ‘zymase’ = retained the ability to convert sugar into alcohol later show conversion result of serious chem reaction by spec enzymes. Demonstrated ‘no vital force’, enzyme activites can be purified and worked on in vitro

Question 9

How is Karly Ereky attributed with the development of the modern biotechnology process?

Answer 9

Coined the term ‘biotechnology’ 1917 process farm pigs w/ sugar beet as food staple 
Sugar beet (raw material) -> prepare foodstuff (upstream processing) -> feed pigs (biotransformation) -> slaughter pigs (downstream processing) -> sell pork (pure product)

Question 10

How did EREKY define biotechnology?

Answer 10

All lines of work by which products are produced from raw materials by living things

Question 11

How was the production of antibiotics optimised to increase yield (progress in 1940s – 70s)

Answer 11

Improved strains of bacteria/fungi used; improved growth conditions ‘submerged culture’ fermentation; mutation and selection to increase yields from microbes

Question 12

What features of microbes make them useful for biotechnology?

Answer 12

Small size and high surface area ratio (facilitate rapid nutrient transport); high metabolic rate; adaption to broad range of environments, diff metabolic processes; grow well on low grade inexpensive + sometimes toxic materials

Question 13

How did COHEN and BOYER develop recombinant DNA technology + what was their experiment?

Answer 13

Met @ conference and combined research ideas – COHEN transferring plasmids between bacteria, BOYER enzyme cut DNA @ specific sites (restriction endonuclease)
1873 cut up plasmid with restriction enzyme + spliced in foreign DNA + transferred products to E.coli
FIRST RECOMBINANT DNA MOLECULE CREATED USING GENE SPLICING

Question 14

Who developed dideoxy sequencing and what is it used for?

Answer 14

Dideoxy sequencing = chain termination method, used in 1980s by Fred SANGER to sequence small regions of DNA

Question 15

What was the first full genome to be sequenced and what and who and when?

Answer 15

Haemophilus influenzae 1.8Mb genome by Craig VENTER + al 1995 by chain termination method
Published E.coli in 1996

Question 16

What other model organisms have been sequenced and when?

Answer 16

Budding yeast (Saccharomyces ceravisae 1996); Arabidopsis thaliana 2000; Human 2001

Question 17

Advantage of development of sequencing methods and 2nd and 3rd gen methods?

Answer 17

Allow sequencing of billions of bp at low cost

Question 18

Who was PCR developed by, when, and what for?

Answer 18

1988 by Kery MULLIS, allowed rapid amplification of DNA pieces for cloning – combined with knowledge of genome sequence allowed genes to be easily clones and manipulated

Question 19

How have genetic methods been improved in microbes such as yeast?

Answer 19

Though the use of viral vectors to intoduced cloned recombinant genes for expression

Question 20

When were Ti (tumour inducing plasmids from certain bacteria) plasmids first utilised and how?

Answer 20

In 1983, to transform plant cells with recombinant plasmids

Question 21

How can Site Directed Mutagenesis improve properties of recombinant proteins?

Answer 21

SDM means that it is easy to design and create mutated copies of a gene to allow for specific changes in protein sequence

Question 22

What does in silico mean?

Answer 22

simulated on a computer

Question 23

What is the benefit of allowing whole cell modelling of metabolism in silico?

Answer 23

allows microbiologists to predict how to manipulate important bacterial properties for biotechnology uses

Question 24

What are two advantages of gene synthesis compared to the use of PCR?

Answer 24

1) Allows for codon optimisation

2) Can make changes to a sequence, e.g. add a restriction site OR make a specific mutation

Question 25

When was the first whole genome synthesised and by whom?

Answer 25

by craig VENTER et al. 1.08Mb Mycoplasma mycoides genome from oligo nucleotides -> used to replace host genome for a particular Mycoplasma species