BIOL%112:%Capstone%topic%on%Bacterial%Cell%Growth%

In#thinking#about#how#a#population#of#bacteria#grows#over#time#in#liquid#culture#media,#this#topic#is#
intended#to#get#students#to#integrate#and#apply#many#of#the#cell#biological#processes#we#have#learned#
so#far#in#BIOL#112.#
#
Learning#objectives:#Bacterial#cell#growth#
! Describe! what! is! meant! by! growth! in! a! unicellular! organism! distinguishing! between! growth! and!
division!of!individual!cells!and!growth!of!a!population!of!cells.!
! Identify! the! four! phases! of! population! growth! in! a! batch! culture! and! in! the! associated! growth!
curve!graph!8!lag!phase,!exponential!phase,!stationary!phase,!and!death!phase.%!
! Compare%and%Contrast!a!population!of!cells!in!lag!phase,!exponential!phase,!stationary!phase,!and!
death!phase!in!terms!of!division!rate,!cell!composition,!and!cell!survival.!
! Predict%the!changes!in!a!population!of!cells!and!the!observed!growth!curve!in!response!to!changes!
in!environmental!conditions.!
%
Pre?reading%for%Bacterial%Cell%Growth:%
!
One! of! the! oldest! life! forms! on! earth,!
bacteria! are! single! celled,! microscopic!
organisms!that!are!extremely!diverse!in!their!
habitats!and!metabolic!needs!(Figure!1:!HLW!
Figure!26.1).!As!we!have!learned!throughout!
this!course,!a!bacterial!cell!can!be!considered!
one! of! the! smallest! units! of! life.! This! single!
cell!is!capable!of!self8replication,!storing!and!
transmitting! genetic! information,! harnessing!
energy! from! the! environment! and!
maintaining!a!different!internal!environment!
Figure 1: HLW FIG. 26.1 A bacterial cell. The cells of
inside!the!cell!separate!from!the!surrounding! Bacteria (and Archaea) do not have a membrane-
environment!outside!the!cell.!! bounded nucleus or other organelles.
! !
There! are! many! diverse! species! of! bacteria! and! different! types! of! bacteria! can! grow! in! different!
environments! such! as! in! soil,! in! water! sources,! and! on! plants.! Some! bacteria! can! even! thrive! in!
extreme! environments! like! the! high! temperatures! found! in! hydrothermal! vents! on! the! ocean! floor.!
Bacteria! are! often! living! symbiotically! with! other! organisms.! In! humans,! many! different! types! of!
bacteria! normally! reside! in! a! number! of! our! tissues! including! our! skin,! reproductive! and! digestive!
tract.! A! healthy! microflora! is! integral! for! normal! human! development! and! physiology.! For! example,!
by8products!produced!by!the!many!communities!of!bacteria!living!within!us!contribute!to!a!normally!
functioning!immune!system!and!digestion!of!our!food.!Bacteria!are!also!utilized!intensively!for!applied!
industrial! applications! including! vaccine! production,! biotechnology! applications,! cleaning! up!
environmental!disasters!like!oil!spills,!and!production!of!fermented!food!products!such!as!yogurt!and!
cheese.!All!of!these!applications,!not!to!mention!the!widespread!use!of!bacteria!as!an!instrumental!
research!model!for!studying!conserved!processes!in!cell!biology,!make!understanding!the!growth!and!
function!of!bacteria!is!of!great!importance!to!sustain!an!amazingly!diverse!number!of!fields.!
!
Bacterial! cells! grow! and! divide! through! a!
process!termed!binary!fission!(Figure!2:!HLW!
Figure! 11.1).! Binary! fission! is! a! form! of!
asexual! reproduction.! The! result! of! binary!
fission!is!the!formation!of!two!identical!cells!
from! one! originating! bacterial! cell.! Different!
bacteria!can!achieve!cell!division!at!different!
rates.!And!even!the!same!bacteria!can!divide!
at! different! rates! depending! on! the!
environment!in!which!it!resides.!For!example,!
the!intestinal!bacteria!Escherichia#coli!(E.#coli)!
will! grow! at! a! much! slower! rate! residing!
inside!our!gut!compared!to!the!rate!at!which!
it!can!grow!under!optimal!conditions!in!a!lab.!
!
Environmental! conditions! such! as! levels! of!
available! nutrients! (and! also! competition!
between! cells! for! cellular! uptake! of!
nutrients!),! temperature,! pH,! and! osmotic!
pressure! all! contribute! to! the! rate! a! cell! can! Figure 2: HLW FIG. 11.1 Binary fission. Cell division in
divide.! As! such! this! will! affect! the! overall! bacteria and archaeons occurs by binary fission.
growth! of! a! whole! population! of! bacteria!
within! a! particular! environment.! Growth! rate! of! a! population! of! bacteria! is! often! expressed! as!
generation%time%(g)!(Figure!3).!Generation!time!is!the!time!it!takes!the!population!of!cells!to!double.!
Generation! time! varies.! Back! to! our! E.# coli! example,! in! the! lab! under! optimal! conditions,! the!
generation!time!for!E.#coli!is!approximately!every!20!minutes,!while!the!population!of!intestinal!E.#coli!
only!double!every!12824!hours.!!
!
Let’s! consider! growing! bacteria! like! E.# coli! in!
the! lab.! When! bacteria! are! placed! into! a!
favourable! growth! environment! with! a! rich!
source!of!all!the!right!nutrients,!they!growth!of!
the! bacterial! population! will! follow! a!
characteristic! pattern.! This! can! be! shown! be!
looking! at! the! number! of! cells! present! in! the!
population! (on! a! logarithmic! scale)! over! time.!
To! do! this! in! the! lab,! the! bacteria! of! interest!
are% inoculated! into! a! sterile! nutrient% media,!
creating! a! bacterial% culture.! This! culture! is!
incubated! at! optimal! growth! conditions!
specific! for! the! bacteria! of! interest! (Figure! 4).! Figure 3: Generation time of a bacterial cell.
Growth! curves! are! created! by! plotting! the! log!
number!of!live!bacterial!cells!present!in!the!culture!(y8axis)!per!unit!time!(x8axis).!Growth!curves!have!
four!distinct!phases:!lag!phase,!exponential!phase,!stationary!phase!and!death!phase!(Figure!5).!
!
!
%
Phases%of%bacterial%growth!
%% A!
Lag%phase.%As!a!student,!when!
you! first! arrived! at! university,!
there!was!no!doubt!a!period!of!
adjustment! as! you! adapted! to!
your! new! surroundings.! Some!
of!you!needed!to!adjust!to!new! B!
roommates,! finding! your!
classes,! developing! skills! to! 1%hour% 2%hours% 3%hours% 4%hours% 5%hours% 6%hours% 7%hours%
study! and! perhaps! you! even!
needed! to! learn! how! to! cook!
for! yourself!! Suddenly,! you!
needed! different! tools! at! your! Figure 4: A single colony of live bacteria growing on a petri dish is
disposal.! Similarly,! when! transferred to the rich nutrient liquid media (panel A). During incubation,
bacteria! are! inoculated! into! a! the liquid media becomes cloudy as the bacterial population grows over time
nutrient! media! they! also! have! (Panel B). (Courtesy of Katelyn Knuff, BIOL 112 TA and PhD Candidate in
Dr. Brett Finlay’s lab, UBC).
a! time! of! adjustment.! During!
lag! phase! individual! bacterial!
cells!are!growing!but!not!readily!dividing.!At!this!phase,!the!bacteria!cells!are!making!the!tools!they!
need!to!thrive!in!their!new!environment!(just!like!those!ear8plugs!allowed!you!to!thrive!with!your!new!
roommate!).! These! tools! involve! expressing! genes! that! encode! for! particular! proteins! (recall! 8! gene!
regulation!and!transcription!and!translation).!These!proteins!include!structural!proteins,!enzymes!that!
break! down! nutrients! available! (recall! the! genes! in! the! lac! operon),! integral! membrane! proteins!
(recall!8!membrane!transport),!or!synthesizing!macromolecular!building!blocks!for!cell!structure!and!
growth! (recall! –! macromolecules).! The! length! of! this! phase! depends! how! different! the! current!
environmental!conditions!were!from!the!last!environment!in!which!the!bacteria!was!found.!While!the!
cells! are! actively! busy! and! growing! in! preparation! for! active! cell! division,! the! overall! population! of!
bacteria!(thinking!about!all!the!cells!present!in!the!media)!does!not!display!a!net!increase!in!growth!at!
this!phase.!!
%
Exponential% Phase.% Imagine! it! is!
half! way! through! your! first! year!
at! university.! You! have! learned!
Log!numbers!of!live!cells!

where! your! classes! are,! how! to!

cook! your! meals! and! how! to!
‘deal! with! your! roommate’.! You!
are! now! fully! adapted! to! your!
environment.! As! a! result,! you!
start! to! make! friends,! find!
favorite!study!areas!and!begin!to!
maximize! all! the! resources!
available! to! you! on! campus.!
Similarly,! when! bacteria! have! Time!
adapted! fully! to! their!
Figure 5: Bacterial growth curve
surroundings!they!will!also!maximize!all!the!benefits!they!can!from!the!nutrient!media,!and!at!optimal!
conditions! bacteria! want! to! replicate!! Exponential! phase! can! be! defined! as! the! phase! where! the!
overall! population! of! bacterial! cells! undergo! binary! fission! at! a! constant! rate.! Replication! is! at! its!
maximum!and!cell!number!increases!exponentially.!One!bacterial!cell,!will!become!2,!which!will!result!
in! 4,8,! 16…etc.! This! can! be! expressed! mathematically! as! 2n! ;! n! being! the! number! of! generations.!
During!this!phase!the!cells!are!readily!dividing!and!the!growth!of!the!population!occurs!exponentially.!
Expressed! on! a! logarithmic! scale,! it! is! easy! to! see! that! the! growth! of! the! population! as! a! whole! is!
occurring!as!at!a!constant!rate!as!evident!by!the!slope!of!the!curve.!!
!
Stationary%Phase.!At!this!phase,!the!key!nutrients!in!the!media!are!beginning!to!deplete;!therefore,!
bacteria!begin!to!compete!with!other!cells!in!order!to!obtain!nutrients.!In!a!lab!culture!like!this,!the!
bacteria!are!also!contained!in!a!closed!vessel.!Metabolic!waste!products!released!from!the!bacterial!
cells!begin!to!build!up!in!the!environment.!This!build!up!of!waste!is!toxic!to!many!of!the!bacterial!cells.!
When! we! observe! this! phase! on! the! growth! curve! we! see! the! curve! plateauing.! This! plateau!
represents! a! net! zero! growth! in! the! overall! population,! which! results! from! an! equal! amount! of!
individual!cells!actively!dividing!as!there!are!cells!that!are!dying.!!
!
Death% Phase.! In! this! stage,! the! loss! of! nutrients,! build! up! of! waste! products! and! lack! of! optimal!
conditions! results! in! the! death! of! many! bacterial! cells.! Overall,! in! the! population! of! bacterial! cells,!
more!cells!are!dying!than!actively!dividing.!This!is!why!we!see!a!drop!in!the!number!of!live!cells!on!the!
growth!curve,!which!means!that!growth!of!the!overall!population!dramatically!decreases.!